CADlock Presentations

Coronary Artery Disease (CAD) is responsible for approximately 17.8 million deaths worldwide annually (Brown et al., 2022). The prevalence of the disease continues to rise in the southern portion of the United States, including the state of Georgia (Norton, 2016). Moreover, it’s a progressive illness as the arteries become blocked with the gradual build-up of plaque. However, these long-term effects are highly preventable through proper diet and exercise. Yet, awareness of these healthy habits is substantially low in the general population, especially among females and people of color (Cushman et al., 2019). As such, Lambert iGEM focused on overcoming this issue by emphasizing the pathophysiology and effects of coronary artery disease, the importance of heart-healthy habits, and CADlock’s role in combating CAD.

Since CAD is a chronic disease, risk factors can develop as early as childhood, eventually leading to the onset of the disease in adulthood. They range from immutable causes like sex, age, familial history, and/or heredity to lifestyle choices, including cigarette smoking, obesity, hypertension, physical activity, and most importantly, diet quality (see Fig. 1) (Nix, 2021). These risk factors are significantly higher in the Southeastern region of the United States, making intervention through effective treatments and preventative methods at an early age even more critical (Casteel, 2014). Lambert iGEM addressed these trends by creating our biosensors (see RCA) and educational materials to encourage proper health and nutrition.

To inform others about our diagnostic tool and provide heart-healthy information, we held “CADlock Presentations” (see Fig. 2). Lambert iGEM started these presentations by bringing attention to the impact of coronary artery disease on the world, and more specifically, in the Southeastern part of the United States. Additionally, we discussed the pathophysiology of the disease, the risk factors and causes involved, and the state of current detection for CAD. We then tied diagnosis into our project, CADlock, summarizing Micro-Q (see Hardware), wet lab (see RCA), human practices (see Human Practices), and education. Following this, we covered tips for prevention by focusing on two essential components of maintaining heart health: lifestyle and diet. For example, we recommended foods to increase in meals, including fruits, vegetables, fat-free milk, lean protein, and unsaturated fats, and foods to decrease in consumption, such as processed and red meats, sugary drinks, and foods high in sodium. We also reviewed the appropriate amounts of exercise adults and children should be getting per week or day.

Alongside our recommendations, we emphasized that moderation is key. No food should be overly consumed or completely cut out, and there is no need to exercise for hours every day. Saturated fats can still be consumed, and rest days can still be taken as long as one stays mindful about these choices.

Due to CAD’s progressive nature, our primary target audience with these presentations were younger students. Implementing hearty-healty habits from a young age decreases the risk of developing coronary artery disease. As such, Lambert iGEM partnered with our school’s healthcare teachers and the HOSA and Science National Honor Society chapters (see Fig. 3). Additionally, we also worked with our school’s special education program and presented CADlock to exceptional science students (see Fig. 3). Furthermore, we presented our project to middle school students at our transformation workshop (see Transformation Workshops in Synbio Education Section).

During each session, we used interactive slide decks and lessons based on the inquiry-based learning approach, and we included a mini quiz-like game at the end. To measure the effectiveness of our lessons, we conducted pre- and post-surveys (see Fig. 5). In total, Lambert iGEM reached over 100 students through our in-person presentations.

Nevertheless, while our focus for CAD education was students, Lambert iGEM also wanted to present heart health knowledge to all cohorts. Therefore, we created a YouTube video and utilized social media and neighborhood group chats to bring attention to it (see Fig. 4). Our content covered the same material as our in-person lessons, making accessibility to this knowledge greater. In total, we amassed over 1000 views on this video and 300 more survey responses. Alongside our presentations, we included a heart-healthy and inexpensive cookbook with numerous recipes from various cultural groups, including within our team, community, and other iGEM teams (see Inclusivity). Through these initiatives, we ensured that the residents of Georgia and elsewhere could access educational material to improve wellness and decrease the impact of coronary artery disease.

Survey Analysis

As shown by Figure 5, at the start of our presentation, over 50% of participants indicated they could not describe the impact of coronary artery disease in the United States, and more than 70% answered that they could not describe how CAD affects the arteries, how the disease is diagnosed, and the importance of miRNAs in detection. Regarding lifestyle and diet, around 50% of those surveyed responded that they could confidently give examples of healthy choices. Interest in the pre-survey on coronary artery disease, miRNAs, and heart health was 27.2%, 35%, and 48.5%, respectively.

By the end of our session, at least three-fourths of respondents stated they could confidently describe the impact of CAD in the U.S., how it affects the arteries and how it is detected, and why miRNAs are crucial for diagnosis. Furthermore, over 97.1% and 95.1% of participants selected they could confidently give examples of heart-healthy habits and foods. Alongside our knowledge survey questions, the interest questions displayed 64.1%, 61.2%, and 67%, respectively, of those surveyed were intrigued by the topics we covered.

Lambert iGEM’s goal to bring awareness to coronary artery disease showed a substantial impact through our CADlock presentations. Overall, our presentations increased understanding and interest in coronary artery disease and our project CADlock. More specifically, in our knowledge questions, we saw confidence increase by 69% in describing the impact of CAD, 69.9% in describing how CAD affects the arteries, 69% in describing how CAD is currently detected, 53.4% in describing miRNAs role in diagnosis, 46.6% in giving examples of heart-healthy habits, and 51.4% in giving examples of heart-healthy foods. Similarly, our interest questions visualized a visible increase in enthusiasm: 36.9% more participants indicated they were intrigued by CAD, 26.2% more participants stated they were extremely interested in learning about miRNAs, and 18.5% showed a high interest in heart healthiness. These differences quantified that those who attended our in-person sessions or watched our video could comprehend the given topics and were engaged. As such, our steps ensured that sound knowledge of coronary artery disease and CADlock is available for the public to access, pushing education forward.

CADlock Presentation References

Synbio Education

Our synthetic biology workshops began in mid-February with our gel electrophoresis workshop. The results of this initiative highlighted the discrepancy in the state curriculum, inspiring us to address the lack of education on biotechnology in Georgia by targeting younger audiences. This resulted in the four major education initiatives we pursued this year in our local counties (see Fig. 1).

Our first initiative was a series of workshops at our local middle school with 7th and 8th-grade students. During these workshops, we ran hands-on experiments that covered DNA structure and function, plasmids and restriction enzymes, and gel electrophoresis.

Our second initiative was a biotechnology summer camp for students from 5th to 9th grade, where students learned about DNA replication, transcription and translation, acids & bases, polymerase chain reaction (PCR), gel electrophoresis, and micropipettes. In both our middle school workshops and summer camp, students reported a substantial increase in their understanding of both the content and interest in synthetic biology after completing the workshop/summer camp, ultimately preparing them to understand synthetic biology concepts in high school.

In August, we offered a transformation workshop to high school students from our previous two initiatives. Students performed a bacterial transformation, utilizing the micropippetting skills they gained alongside the background information they learned. These workshops enabled students to use the knowledge and skills they gained to truly immerse themselves in synthetic biology.

Finally, we hosted a workshop for biology teachers in our state to perform multi-step, inquiry-based lab experiments that were consistent with the 2022 Science Georgia Standards of Excellence. Teachers who attended this session learned about genetics and biotechnology labs and their corresponding activities, so they could enhance their own labs for their students, helping increase our outreach and guaranteeing the longevity of synthetic biology education.

Alongside our four major initiatives in synthetic biology education, we also worked closely with a senior living home to perform a DNA extraction, including the eldery in conversations about science.

Throughout every initiative, we used a scaffolded and inquiry-based learning approach to increase engagement in synthetic biology (Pedaste et al., 2015). Ultimately, teaching synthetic biology education across the community not only ensures that the biological foundation is built for students but also builds their future in an expanding field.

Framework

In each of our 2022 education initiatives, we followed an inquiry-based, scaffolded teaching approach (see Fig. 2). By combining both strategies, Lambert iGEM built a foundation to spread fundamental scientific background, and later, synthetic biology to students in younger grade levels.

In inquiry-based teaching, students can observe and ponder phenomena, create explanations, conduct experiments and gather data, and support theories through a hands-on learning experience. Furthermore, it allows them to understand and evaluate why certain situations or results occurred. This type of teaching has been proven to decrease teacher-led discussions and improve critical thinking skills in students (Pedaste et al., 2015). Lambert iGEM applied the ideas of inquiry-based teaching in each of its events hosted this year. For instance, students at the Riverwatch workshops were introduced to gel electrophoresis; after a brief explanation, students were able to understand a protocol and utilize gel electrophoresis in a common application: forensics. After completing the lab, each student reflected on what worked, what did not, how it happened, and other possible applications. This approach allowed students to grasp the new concepts and develop their knowledge in synthetic biology, as determined by our post-surveys (For further description, see Riverwatch Workshops).

To support students throughout our various events and develop a strong foundation for synthetic biology in students, we connected our inquiry-based workshops with a scaffolded learning approach. Scaffolded learning is a technique that assists learners as they move from one skill to the next and gain achievements at each level (Maybin et al., 1992). The interconnectedness of this method allows students to transition between difficult levels with a comprehensive understanding of previous material. Students will have the scientific background knowledge to succeed in later workshops. Additionally, utilizing a scaffolding approach maximizes the effectiveness of inquiry-based learning, as this method allows students to focus their attention on relevant features and increase their ability to learn advanced concepts. The combination of the two techniques enhances problem-solving, reflection, research assistance, concept integration, and knowledge acquisition skills (Pedaste et al., 2015). As a result, Lambert iGEM incorporated both inquiry-based and scaffolded learning by hosting a multitude of events strategically placed throughout the year. Later in the fall, the same students from the previous workshops and camps attended the transformation workshop in which we presented our project: CADlock. As a result of scaffolded learning throughout the year, students were easily able to succeed in the lab, as seen by the survey data in this section (see Transformation Workshops). By laying strong groundwork earlier on and including new information at each lecture, while still ensuring understanding of previous content, scaffolding proved effective.

Overall, Lambert iGEM’s approach to education and spreading synthetic biology this year combined two complementary techniques: inquiry-based teaching and scaffolded learning. They allowed students to fully grasp each concept and take these new skills to succeed in the next “stages.” The implementation of such techniques in each of our hands-on workshops proved favorable. Each of the events mentioned above is described in further detail below to demonstrate how both techniques were used and their efficacy.

Gel Electrophoresis

In February 2022, Lambert iGEM hosted a Gel Electrophoresis Lab for freshmen at our school. These students were unable to perform gel electrophoresis in the biotechnology unit of their biology classes even though this topic is a key component to understanding biotechnology. Therefore, we invited approximately 20 students to participate in this workshop and gain hands-on experience.

We split this lab into 3 sections: fundamental skills (ex: micropipetting); gel electrophoresis background information, purpose, and applications; and the lab itself, where we tied in an interactive forensic story for the students to follow. Throughout the lab, we utilized a hands-on approach as students were able to create their own gels, pipette the reagents into the wells, and run the gel electrophoresis themselves.

While we did not conduct pre and post-surveys, students showed considerable engagement in this topic and enjoyed the experience of the lab. Throughout and after the lab, students mentioned how they wished they could explore concepts in biotechnology further, and how the biology courses they took didn’t adequately prepare them for more application-styled units such as biotechnology. For instance, many students stated that before our Gel Electrophoresis Lab, they struggled with concepts like DNA and protein structure-function, two key topics addressed in the workshop. Even though we recognized the lack of biotechnology in the curriculum of classes like 9th grade honors biology, the Gel Electrophoresis Lab brought our attention to gaps rooted in the state science curriculum being taught in middle schools. Consequently, we reviewed the Georgia Standards of Excellence for Science and found that the content only covered the involvement of genes and chromosomes in inheritance, leaving off fundamental topics including DNA and protein structure/function (see Fig. 5).

The results of the Gel Electrophoresis Lab inspired us to address this absence by introducing these concepts to students in earlier grades. Furthermore, by creating more quantitative sources of feedback, such as surveys, we were able to adapt our curriculum to the needs of the students based on their responses. Thus, the Gel Electrophoresis Lab led us to utilize the scaffolded and inquiry-based learning approaches to target younger audiences, leading to our first major initiative: the Riverwatch Middle School workshops.

Riverwatch Middle School Workshops

Lambert iGEM hosted a series of six workshops at Riverwatch Middle School to increase awareness and engagement around synthetic biology and address the lack of it in the Forsyth County school system at the middle school grade levels. We held two sessions, one for eighth graders and another for seventh, with three workshops for each grade level. These hands-on workshops for middle-school students provided foundational biological knowledge, as we included topics such as proper DNA and protein structure/function and labs such as gel electrophoresis (see Fig. 6).

We organized these workshops weekly throughout the last three weeks of March and the beginning of April. For each workshop, we carefully analyzed which Science Georgia Standards of Excellence for middle school would correspond to fundamental synthetic biology concepts and compiled illustrative slideshows. We reasoned that the biology standards that the students would have mastered in regular schooling would be of more interest to them since they would now be able to apply this knowledge to a different field of science: synthetic biology. In addition, we included a weekly pre- and post-survey to record changes in content knowledge and engagement at each workshop (Nuhfer & Knipp, 2003).

The first workshop covered DNA extraction, re-introducing the structure, function, and purpose of DNA on a molecular and macro level using real-world examples. Then, we completed a wheat germ DNA extraction lab from the Museum of Science so the middle schoolers could physically see and apply the DNA concept they learned (Museum of Science, 2016). In this procedure, students used alcohol and wheat germ liquid to extract clumps of wheat germ DNA by combining them in a tube and inverting it. By explaining the purpose of each material in the lab, the students not only learned about the overall importance of the lab but also gained knowledge about the materials’ purposes; for example, the lab used meat tenderizer, which contains proteases, enzymes that break down proteins. Subsequently, we connected the use of meat tenderizers to the basics of enzymes. After the students completed the lab, we extended the application of DNA extraction to real-world examples: creating sustainable detergents, synthetic food flavoring, etc.

The second workshop focused on plasmids and restriction enzymes. We first explained a plasmid’s structure, location, and purpose, one of synthetic biology’s most widely used molecules. Since not all plasmids perform scientists’ desired functions, we addressed this problem as a question to the students to enable engagement and provide a meaningful transition into restriction enzymes. We explained their purpose regarding plasmids and their importance in our iGEM team. Next, the students completed a hands-on activity to apply all of these concepts.

The students received a strip of DNA base-pairing “letters” and cut out the standardized parts from Biobrick assembly with restriction enzymes on both the plasmid and the DNA to create a new paper circle with the restriction enzyme sites (see Fig. 7 and Fig. 8). This hands-on activity with incremental demonstrations provided the students with the ability to apply these concepts through synthetic biology and see the versatility of DNA.

The last workshop covered gel electrophoresis and basic micro-pipetting techniques. We used a different approach in teaching basic micro-pipetting techniques: we taught the students while demonstrating the concept rather than using a slideshow. We emphasized several crucial points such as the position to hold a micropipette, the importance of changing tips, and the difference between the first and second stops. We then explained the purpose of gel electrophoresis, how micro-pipetting is essential in retrieving accurate readings on a gel run, and how to read gels. To further engage our audience, we created a short forensic prompt to demonstrate how the field can use gels as tools of evidence against suspects in a crime. Upon completing the lab, all the middle schoolers could identify the correct suspect.

As shown in Figure 9, during the first week, over 60% of the students were not very confident in describing why scientists extract DNA. Additionally, approximately 40% of the students ranked their interest as a 3 or 4 in learning about synthetic biology and how DNA can be used. However, by the end of the week, around 90% of the students could confidently and accurately describe why scientists extract DNA; moreover, about 77% and 86% were extremely interested in learning about synthetic biology and the uses of DNA extraction, respectively.

During the second week, only 6.2% of the students were very confident in accurately describing the function and purpose of a plasmid, and 9.4% in describing the function of a restriction enzyme. However, by the end of the week’s workshops, around 43% and 58% of the students were extremely confident in accurately describing plasmids and restriction enzymes, respectively.

At the beginning of the final week, around 20% of the students did not know how to properly use a micropipette and exactly 75% of the students could not describe the purpose of gel electrophoresis in differentiating DNA samples. Moreover, only around 58% of the students demonstrated complete interest in using gels in forensics. However, by the end of the week, 100% and 67% of the students were very confident in describing the function of a micropipette and gel electrophoresis, respectively. Moreover, 82% of the students showed full interest in using gels in forensics.

All of the post-survey pie charts had lower percentages for “no understanding and could not explain” and higher percentages for “understood and might be able to explain” and “understood and can explain” (see Fig. 9). In the post-survey bar graphs, the percentages for all the students who ranked their interest as “5” were substnatially higher than the corresponding percentages in the pre-survey (see Fig. 9). These differences indicate that the students could comprehend the given topics and were engaged in the workshops (Nuhfer & Knipp, 2003). Moreover, the post-survey results also revealed that many students were interested in pursuing synthetic biology in high school and beyond (Nuhfer & Knipp, 2003).

Following the success of the Riverwatch Middle School workshops, we expanded the spread of synthetic biology knowledge and interest from only middle-schoolers to late elementary through early high schoolers in our in-person and virtual summer camps. These camps provided an intensive and organized teaching system of scaffolded learning.

MiniPrep Summer Camps

During the summer of 2022, Lambert iGEM held two introductory biotechnology summer camps for 65 middle school and early high school students (see Fig. 10). Much like other educational initiatives, we focused on inquiry-based learning and a variety of teaching techniques including instruction, repetition, feedback, and explanation (Verenikina, 2008). Our first camp was a four-day, in-person camp held during the first week of June, and our second camp was a virtual camp held on alternating days for 2 weeks during mid-June. We tailored our curriculum to supplement the Science Georgia Standards of Excellence (GSE), which prepared students for classes in high school biology and biotechnology (Georgia Science Standards, 2016).

In-Person Camps

At our in-person camps held in early June, we had students ranging from rising 7th graders (age 12) to rising 10th graders (age 15). For this reason, we divided our students into two age groups: our first group, the Acids, primarily consisted of rising 6th and 7th graders, while our second group, the Bases, consisted of rising 8th to 10th graders. In Georgia, since rising 7th graders have not yet taken a life science class, we were also able to adapt our teaching based on their existing knowledge.

During each day of the camp, we discussed foundational information and allowed students to perform an activity. By doing this, we enabled them to understand the basics of a topic and then build their understanding through experimentation with the materials. The camp’s lab groups consisted of four to five students for a total of about eight groups per room. Each group within the Acids or Bases rooms had one student counselor from our iGEM team, and every room had one designated adult advisor for supervision. We ensured a low student-to-counselor ratio to stress the inquiry-based learning approach. In other words, students could always ask questions to a counselor who would be able to answer them and reinforce their learning.

Day 1

During the first day, we discussed safety considerations throughout the camp: we provided students with goggles, which ensured safety and built good PPE habits for future lab work. Then, we performed two main activities: a DNA extraction and an exploration of acids and bases. For our DNA extraction, we explained the structure and function of DNA using foam pieces that each represented a nucleotide. Then, we followed the protocols developed by the NISE Network’sBuilding with Biology project to extract DNA from wheat germ (NISE Network). The second activity was an acids and bases activity developed by Lambert iGEM. The butterfly pea flower (Clitoria ternatea) can act as a pH indicator, so we made butterfly pea tea for students to qualitatively measure the pH of various solutions, including baking soda, orange juice, and a 10% solution of bleach (Saptarini et al., 2015). Finally, we ended the day by teaching microscope skills. Students swabbed their cheek cells onto glass slides and learned how to focus the microscope.

Day 2

During the second day, we focused on metric conversion skills and micropipetting techniques. We first began with a review of the metric conversion chart, using the mnemonic device: King (Kilo) Henry (Hecto) Died (Deka) By (Base Unit Drinking (Deci) Chocolate (Centi) Milk (Milli). Then, we transitioned into learning micropipetting skills. We tied this into concentration calculations and serial dilutions afterward, which was used for the next lab: Pipetting a Rainbow activity.

In correspondence with our approach to teaching micropipetting at the middle school workshops, we emphasized how to hold a micropipette, the purpose of changing tips, and how to differentiate between the “first stop” and “second stop.” Students applied their new-found skills in micropipetting by mixing small amounts of food colorings to complete the Pipetting a Rainbow activity, creating a color wheel on a coffee filter (see Fig. 11).

We ended the third day by letting the students learn how to use and look through pre-prepared slides with a microscope. We then sent students home with a frugal microscope, called a foldscope, that was purchased from Foldscope Instruments.

Day 3

During the third day of our in person camp, Lambert iGEM gave a lesson over cell membranes to the bootcamp students. We started out with an introduction to cell membranes, a very important biological structure. Cell membranes separate the inside of cells from the external environment, controlling the entrance and exit of nutrients, waste, and other molecules. However, we adapted our lectures and activities on this day to fit the background of our students. For the Acids Group, we brought the students outside to draw cell membranes with chalk on the concrete (see Fig. 12). With our Bases Group, on the other hand, we created an activity where they modeled out cell membranes with Play-Doh (see Fig. 13).

By allowing the younger students to draw out the structures, they could visualize the images from our PowerPoint themselves (see Fig. 15 - Day 3 Slides (6th-7th)). This helped these campers understand the basic function of a cell membrane better. In our older group, by utilizing Play-Doh, they were able to model the cell membrane in detail. As such, they could include more complex components, and build upon surface level material they have previously learned.

Furthermore, by creating lessons based on age groups, the lab that followed this activity was suited to each group’s needs. For both groups, we described cell transport and how that relates to cell membranes through a Gummy Bear lab, explaining how solutes like to move from high concentrations to low concentrations and water likes to move from areas with more water (less solute) to areas with less water (more solute). Nonetheless, we went further in detail in our Bases Group, discussing the hyper-, hypo, and isotonicity of the solutions needed for the gummy bear lab, while we left this off for the Acids Group.

Finally, Day 3 ended with preparation for the PTC lab, where each student received a tube of extraction buffer and was told to follow the cheek cell extraction protocol. Similarly, we explored and explained the concepts differently to different groups. For the Acids Group, we described Polymerase Chain Reaction (PCR) as a means to increase the amount of DNA, while in the Bases Group, we went into full detail behind the process of PCR. Likewise, our reasoning to adapt the lessons to age group were the same as those mentioned above.

Day 4

At the end of the In-Person camp, we picked up where we had left the day before and ran a gel electrophoresis. We explained to both the Acids and Bases group how gel electrophoresis works, what its purpose and application is, and how we were going to use it for the lab. This lab made students draw on the content they learned earlier in the week, including micro-pipetting and dilutions, to pipette the solutions into the gel wells and make the solutions themselves. When the gel finished running, students analyzed the lab to see if they were “tasters” or not, building on genetics that were covered in middle school courses. More specifically, students were taught about being homozygous dominant or recessive or heterozygous. This was then connected to the PTC labs itself, where we discussed that the PTC taster gene is dominant, and those with the genotype of TT or Tt can taste, while those with the tt genotype cannot.

Overall, these campers followed each hands-on lab experience with the help of direct instruction from advisors and counselors, background information, lab protocols, and Q&As. This method of scaffolded learning encouraged student involvement and discussion and allowed for a deeper understanding of their future science curriculum, per the Georgia Education Standards.

Virtual Camps

For virtual camp preparation, we assembled experiments into boxes called the MiniPrep kits. We first planned each kit’s contents by including seven fundamental topics (modules) to design a comprehensive lesson plan for our students. This process included drafting educational lab protocols. We held a test day, where middle school students tried out the experiments. We were able to observe how students of our target age group understood and engaged with the experiment so that we could improve our models. Following these trials, we edited the lesson plans according to where the students struggled with and the parts they found the most interesting. For instance, we originally created a foldable activity for protein structure, but we ended up removing it as students showed little enthusiasm in the activity. Since we targeted a younger audience for the camps, these revisions generally included simplifying background information, troubleshooting the instructions, including more step-by-step guidance within the worksheets, and providing real-world interpretations of lab results so the campers could understand the material.

"Next, we created an inventory of necessary lab materials and packaged each experiment to prepare the kits. Additionally, we created innovative and frugal versions of some of the materials in our labs. These included a home-friendly and cost-effective 3D printed centrifuge used in our DNA extraction lab protocol and a frugal gel electrophoresis kit that ran using batteries, two wires, and an agar gel. These materials contribute to the outreach of our MiniPrep camps and provide the opportunity for a larger population to access biotechnology equipment and protocols by lowering the cost of the kit and making typically expensive lab procedures viable in a cost-effective home setting.

List of Materials The following is a partial list of materials that feature the common supplies the MiniPrep Kit includes across all modules:

• 6 150 mL Beakers
• 5 Pipettes
• 3 250 mL Beakers
• 2 Spoons
• 2 Toothpicks
• 1 Pair of Goggles
• 1 Pair of Gloves
• 1 Syringe
• 1 Graduated Cylinder
• 1 Ruler
• Pen

Along with the general materials listed above, each module had a bag of specific materials needed for experimentation. For example, some of the materials are a baby potato, a 3D frugal centrifuge, a foldscope kit, TAE buffers, gummy bears, butterfly pea tea flowers, food coloring, pepper, PCR tubes, prepared slides, agarose, 3D printed frugal centrifuge, and frugal gel electrophoresis kit.

Additionally, we provided virtual students with a MiniPrep modules packet that contained background information, safety instructions, protocols, and pre-and post-lab questions for each experiment. The MiniPrep modules packet included direct lessons and protocols relating to properties of water, acids and bases, cell transport, enzyme discovery, DNA structure and function, Foldscopes: diversity of cells and life, and gel electrophoresis. We also provided a reference sheet to the students for a better understanding of concepts taught during camp. This sheet featured general background information and helpful aids such as a pH scale, a central dogma diagram, and a macromolecule chart.

Prior to camp sessions, our team also developed MiniPrep PowerPoints, which contained the background information and protocols for each module and incorporated the improvements from our test day. After initial camp preparations, we developed a website for camp registration and resource accessibility, Zoom meeting sessions, an archive of the virtual camp session recordings, and flyers to market our program. We advertised the MiniPrep Camp and encouraged registration through Instagram (@synbiostemed), educators and parents within our county and school system, and the new SynbioSTEMEdu website. (see Fig. 18).

During the virtual camp, students attended Zoom Meetings and participated in experiments with the Lambert iGEM camp counselors. Students also answered pre- and post-lab questions and CER statements (claim, evidence, and reasoning) to reinforce their knowledge and become familiar with basic lab practices. The students read through background information, completed the pre-lab questions, and performed the initial stages of each experiment before the one-hour Zoom meetings three times a week. Then, while in session with their assigned iGEM counselors and advisors, students had the opportunity to review pre-lab material, experiment with a step-by-step walkthrough of each module, and evaluate their results in a post-lab discussion. We set up this schedule to encourage students to work through the experiment cycle independently, while also simulating an inquiry-based learning experience through group discussions and Q&As.

Survey Analysis

Day 1 Surveys

Day 2 Surveys

Day 3 Surveys

Day 4 Surveys

Following these camp experiences, our team offered a pre-and post-survey directly to the students to analyze how effective our lessons and modules were and maximize the amount of content for future students. This survey was administered to our campers, assessing their knowledge, confidence, and overall camp experience. From this data, we found that after attending the MiniPrep camp, students felt definitively more confident in their understanding of these biology concepts. For instance, students who attended the May 31st camp session expressed an average 47.71% increase in the understanding of questions regarding lab setup and biological concepts. Students who participated in the June 1st camp session showed an average 57.57% increase in understanding. And students who participated in the June 2nd camp session showed an average 55.17% increase in understanding. Furthermore, student interest in synthetic biology and lab work increased from the original levels reported before the camp. The data shown in the bar graphs below measure the average confidence levels of students before and after each In-Person day for each of the 5 questions shown in Figure 21. This data from student surveys helped illustrate the effectiveness of the MiniPrep camp in that it not only reinforced knowledge of lab skills and standard biology topics but also that it raised interest and enthusiasm for biotechnology overall. The above-mentioned trends are illustrative of the effectiveness of the scaffolded learning approach that our team focused on throughout the year.

Throughout the in-person and virtual camp experiences, students gained vital skills for experimentation, explored an introduction to high school biology topics such as central dogma and acids and bases, and worked through various biotechnology processes such as gel electrophoresis and DNA extraction. By holding these camps, we empowered students with an immersive experience of foundational biology and chemistry. These skills could be utilized in future iGEM workshops like the transformation workshop. All in all, our summer camp initiatives ensured that students grasped these fundamental concepts and understood the variety of real-world applications synthetic biology provided.

After receiving positive results and feedback from our summer camps, we continued the spread of synthetic biology by inviting these students to our Transformation Workshop. This lab empowered students to execute standard bacteria transformation, examine transformation efficiency, and use common lab equipment such as micropipettes. This final workshop provided an efficient way to tie together concepts learned at the middle school workshops and the summer camp to synthetic biology, allowing students to fully immerse themselves in this field.

Transformation Workshop

As the final piece of our 2022 scaffolded learning approach, Lambert iGEM designed a transformation workshop, adapted from BioBuilder, targeting the same demographic and developing the synthetic biology techniques students learned at the RMS workshops and MiniPrep summer camps. The lab involved transforming bacteria to fluoresce different colors, allowing the students to easily observe their transformation efficiency. Moreover, it allowed the students to apply their micropipetting skills in a practical setting, advance their understanding of basic genetic properties and processes, and be introduced to synthetic biology at an earlier age.

We utilized our social media accounts and created illustrative fliers to bring interest to this final initiative, as our target audience was freshmen and sophomores at Lambert High School who attended our prior two workshops.

We started planning at the beginning of July, and the workshops lasted three days in the last week of August. To structure the program and make the concepts more easily digestible for the participants, Lambert iGEM created slideshows and activities to complement each day. Additionally, Lambert iGEM created a pre-survey for students to fill out on the first day, so we could understand how initially well-versed students were in synthetic biology and bacterial transformation.

Day 1

On the first day, students learned about plasmids, their distinction from chromosomal DNA, and their application in genetically transforming bacteria. We did this by describing horizontal gene transfer in a natural setting. After the plasmid lecture, students were given plasmid manipulatives, where they were able to swap out different parts of a plasmid, including origins of replication, antibiotic resistance genes, and gene inserts. They were also able to “transform” these plasmids into two different strains of bacteria, a protein expression strain, and a cloning strain. Then, they could “plate” these bacteria onto plates, which were either plain or had different antibiotics. Throughout this activity, students were asked to think about the type of colonies they could expect to see with different ORIs, antibiotic resistance genes, insert genes, bacteria, and plates (see Fig. 25).

Finally, students were introduced to the two different chassis and signaling pathways used in the lab and the description of each: the BL21 E.coli was designed for increased protein expression, while the K12 strain was made for cloning; moreover, for the two plasmids used, pPRL is a longer signaling pathway and requires more resources, while pGRN is shorter and requires less. Following this, we asked the students questions about why the difference in chassis could be significant, especially regarding the different signaling pathways.

Day 2

During the second day, students used the given procedures to perform the lab and gather data. Students were provided with microcentrifuge tubes, Luria broth, CaCl2, six plates (five with ampicillin and one plain plate), ice, a sterile inoculating loop, micropipette and pipette tips, latex gloves, waste beakers, and sharpies. Four microcentrifuge tubes were labeled 4-1 pGRN, 4-2 pGRN, 4-1 pPRL, and 4-2 pPRL (4-1 is strain K12, and 4-2 is strain BL21). Next, students transformed both strains of bacteria with the provided plasmids in the respective tubes and spread 250 microliters of each solution onto four labeled agar plates which were incubated overnight.

Day 3

On the final day of the workshop, students examined the growth on their four plates. While there were varying results (likely due to errors in micropipetting and heat shocking), results generally showed a relatively large amount of growth on the 4-1 pGRN plates, less but darker green growth on 4-2 pGRN, a few small purple colonies on 4-1 pPRL, and no growth on 4-2 pPRL. After giving ample time for students to study their own results and go around the room to observe and compare data, they were given post-lab questions to promote critical thinking and utilize the concepts of limited cell resources and optimization of differing chassis.

Survey Analysis

At the end of the third day, we asked participants to fill out a post-survey to provide feedback on the quality of the workshops and the clarity of the material taught in comparison to their responses on the pre-survey.

The data collected shows a large increase in confidence in synthetic biology and the process of transforming bacteria. For instance, in the pre-survey, most, or all, students were unconfident, if not completely new, to the topics taught throughout the workshops but demonstrated great improvements, as observed in the post-survey. Additionally, participants were asked about their further interest in specific topics covered throughout these three days. There was a considerable demonstration of interest, indicating students were invigorated and would likely pursue further knowledge in synthetic biology.

Beginning at Riverwatch Middle School, Lambert iGEM addressed the gap in the middle school science standards. We started by introducing topics like DNA structure, function, and replication as well as transcription and translation. Then, we discussed restriction enzymes, gel electrophoresis, and techniques like micropipetting. Through this scaffolded learning approach, by the time these students entered the Transformation workshop, they had all the background knowledge necessary to understand the lab. Thus, by the end of this final workshop, the majority of students were fully prepared to learn and understand synthetic biology, illustrating the success of our 2022 Education Initiatives.

Teacher PTC Workshops

Since our previous three initiatives set a standard for teaching synthetic biology to younger students, we sought to increase our impact and provide this content to students across different age groups in both the county and the state.

Therefore, Lambert iGEM hosted a workshop for teachers around our county, utilizing the Phenylthiocarbamide (PTC) Lab adapted from miniPCR bio as a vector, providing hands-on experience and empowering teachers to integrate synthetic biology into their curriculum. This workshop allowed teachers to use instruments such as micropipettes, thermocyclers, and gel electrophoresis. Later, they interpreted the gels using the knowledge we covered during the lab to examine their cheek cells and test for the PTC tasting gene. Finally, we tied a chi-square and Hardy-Weinberg activity with the lab to see if the sample of taste testers for PTC matched the general populations, giving teachers a means to teach a complex topic.

By the end of the workshop, teachers were equipped with the necessary tools to bring the activities and experiments to their own students, ranging from 6th-grade to 12th-grade students.

Lambert iGEM planned the workshop throughout the summer and held it during the third week of August. The workshop covered high-school biology standards from the Science Georgia Standards of Excellence (Georgia Science Standards, 2016). By creating flyers (see Fig. 25) and partnering with Dr. Brittney Cantrell, Secondary Science Content Specialist for grades 6-12 in Forsyth County, we advertised our idea to teachers around our county and acquired a strong audience to participate in this lab.

PTC Taster Lab Protocols:

1. Collect cheek cells by gently scraping the inside of their cheeks with the flat end of the provided toothpick.
2. Swirl around the toothpick in the microcentrifuge tube containing 50uL of extraction buffer to transfer the cells into the buffer.
3. Incubate the tube for 10 minutes at 95°C in the incubator
4. Pipette 12.5 uL of master mix, 12.5 uL of primers, and 3uL of cheek cell mixture into the PCR tube.
5. Place the PCR tube into the thermocycler to amplify the PTC taster gene using PCR.
6. To make the gel, mix 0.5g of agarose powder and 50mL of TAE buffer provided.
7. Microwave the mixture until it appears clear.
8. Pour the liquid into the well box containing a well comb and let it cool until the gel has hardened.
9. Pipette 12uL of DNA Ladder into one well
10. Pipette 15uL of cheek cell DNA into another well.

When the teachers arrived, we provided them with a PowerPoint (see Fig. 33) that they would use to teach their students about the lab. They also received a folder containing the teacher handbook (see Fig. 34) to provide supplementary information about the lab. To provide an overview of the topics covered in the lab, we talked about iGEM, synthetic biology, and genetics. Then, they completed a cheek cell extraction using many of the tools and devices used in professional labs to apply the content. After incubation and PCR, the teachers made their gel and practiced pipetting into practice gels while they were letting the experimental gel cool. Once they pipetted their cheek cell DNA and started gel electrophoresis, we demonstrated how chi-square and Hardy-Weinberg problems could be correlated to this lab to include another way the science standards could be embedded into this lab (see Fig. 35). After analyzing the gel results, we summarized the ways in which this experiment is crucial to introducing synthetic biology to the younger generation and how teachers can use the provided information to teach their students about such a thriving field of science.

Results

To receive feedback, we collected data via a form from participants regarding their overall enjoyment and the potential ways they were planning on implementing the workshops in their classrooms. The workshop ran successfully as results and feedback showed a large increase in interest and knowledge. Teachers were confident in their skills as they experienced an important lab in synthetic biology. Since they worked with many different professional tools (see Fig. 36), they were opened up to a new field of science that they wanted to incorporate into their syllabus for their students to learn. In addition to the workshop, we presented our CADlock presentation and received both positive feedback and advice on improvements to the project.

Figure 36. Teachers from the workshop practiced pipetting water onto micropipette practice cards

Through the positive results and feedback we received from the teacher PTC workshop, we achieved our ultimate goal of shaping DNA technology, biotechnology training, and activities available in an informative and interesting way. Moreover, PTC taste strips, an incubator, a thermocycler, and gel electrophoresis not only effectively implement biotechnology in the experiment, but cover the use of many of the main devices in the field. Since this workshop did not require extensive research and long working hours, the teachers participating in the lab found it engaging, motivating them to bring this lab into their science classroom curriculum. Additionally, because the teachers were able to work with real DNA and use it in gel electrophoresis, the hands-on activities further validate the lab’s importance toward this audience group as it enables students to share the same experiences.

Legislation

To ensure the longevity of synthetic biology education, Lambert iGEM plans to develop legislation to supplement state-wide curriculum with 3D lessons related to synthetic biology. Our activities would provide opportunities for students to immerse themselves in a novel field by obtaining evidence, evaluating results, and communicating goals with others to further promote science literacy. In October, we reached out to the Georgia Science Program Specialist, Dr. Shirley-Stevens, of the Georgia Department of Education (GaDOE) regarding the development of such legislation. She informed us that the district level largely handles curriculum decisions, which led us to contact the District Second Science Specialist, Dr. Brittney Cantrell.

Dr. Cantrell helped us understand the differentiation between state and district level curriculum. The GaDOE sets the minimum standards for education, which the district level can expand on to provide more focused experiences. For our lessons, we would have to develop about 3-4 class periods of material, with fully fleshed-out lesson plans including background materials, teacher preparation, student protocols, and purchasing lists.

In addition, she advised us on the timeline and barriers to implementing the activities that we proposed. One potential timeline would be to pilot this program during the spring of 2023, with the hopes of district-wide implementation during the 2023-24 school year. Another option would be to hold a workshop during the teacher development days in October, January, and February including the material we planned to implement in our lessons. Both of these possibilities allow for time to receive feedback from teachers to improve our content. In addition, we may be able to implement these at other state-wide teacher development organizations, since introducing this at a state level would be difficult.

Figure 37. Discussions with the Georgia Science Program Specialist (Dr. Shirley-Stevens) and District Second Science Specialist (Dr. Brittney Cantrell) about supplementing the state curriculum.

Antebellum

The hope to spread scientific interest and synthetic biology to all ages inspired Lambert iGEM’s Antebellum initiative. As we worked with students and teachers in our community, we became aware of the possibility of including older adults in our scientific conversations, an initiative that will not only promote brain health through stimulating activity, but also allow our team to become more involved in our community (NIA, 2020). Through Lambert High School’s HOSA chapter’s pre-existing relationship with Antebellum, a senior long-term healthcare facility, we sent email correspondence to Mrs. Neha Mulkar, who arranged our Antebellum workshop. On September 22, we hosted a wheat germ DNA extraction workshop (see Fig. 38). During this workshop, we conversed with residents, helped out during their experimentation, and shared their excitement in performing the DNA extraction. This workshop not only provided us with the experience to share our passion with the residents, but also expanded our views on how synthetic biology can be integrated into our community. In future years, we hope to continue our workshop at Antebellum and slowly introduce similar workshops at other local long-term living facilities.

Figure 38. Sam (Left) and Ricky (Right) helping Antebellum residents throughout the lab.

Conclusion and Future Direction

Through our programs, we addressed the lack of synthetic biology standards and education in the Forsyth County School system and beyond. The engagement of rising middle through high school students and science teachers in our educational workshops, camps, and other activities has contributed to a stronger foundational understanding of biology, leading to engagement in synthetic biology.

Our approach of using inquiry-based and scaffolded learning in our programs expanded the overall understanding of science concepts by allowing students to develop critical thinking, knowledge acquisition, and problem-solving abilities. By implementing these methods, we found that our students almost always gained new integral biotechnology knowledge, as seen by over 50% improvement rates in almost all of our pre- and post-surveys, from the Riverwatch to the Transformation workshop.

Our future goals include:

Figure 39. Our 2023 education goals.

Our goals for 2023 include working closely with our county and state department’s of education to supplement classrooms with 3D lessons in synthetic biology. As such, we can not only address the lack of molecular biological concepts in DNA and protein structure and function at its core, but we can also ensure our impact in synthetic biology education reaches hundreds of thousands of students. To facilitate these curriculums, making them easier for teachers to implement, we plan on developing handbooks with pre- and post-labs, CERs, and interactive instructions and discussion for teachers to use. Furthermore, Lambert iGEM aspires to create grade-specific material alongside the handbooks, allowing students to understand this content in a manner that is digestible in each age group. Finally, we hope to expand our educational outreach beyond Georgia to an international audience with activities adapted to various languages. By reflecting on our work, progress, and feedback from the workshops this year, Lambert iGEM hopes to improve these programs to continue to educate and inspire the future of biotechnology.

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