Attributions

The increase in local and global prevalence and deaths due to coronary artery disease (CAD) inspired Lambert iGEM’s 2022 project, CADlock. Since current CAD detection methods are invasive and expensive, our team researched padlock probes and microRNAs (miRNA) and utilized frugal hardware components to develop Micro-Q, a fluorometer based on light intensity. Additionally, Lambert iGEM created a software device, Probebuilder, that simplifies the process of designing padlock probes.

	Hardware	Software	Wiki	Model	RCA	RCT	Characterization	Implementation	IHP	Education	Scripts	Entrepreneurship	Graphics	Video	Inclusivity	Collaboration
Srijyoth Ashok
Sishnukeshav Balamurali
Varnica Basavaraj
Akhilesh Bharatham
Christina Cho
Ryan Du
Manaswi Gorle
Madhav Gulati
Manasvi Gupta
Richard Jiang
Michelle Jing
Daeun Lee
Nicholas Lockhart
Hari Mudigonda
Rathi Murugan
Sahana Narayanan
Suhayb Nijim
Hannah Noh
Sihyeon Park
Akshaya Poonepalle
Vineeth Sendilraj
Natalie Tan
Srirag Tatavarti
Shivaek Venkateswaran
Vandana Vijay Kumar
Sam Yeleti
Christina Yi
Rick Zhao

miRNA Researchers

Dr. Robert Meller is a cardiologist bench researcher who has been partaking in research regarding RNA biomarkers at Morehouse Medical Hospital. He reviewed the fundamentals of RNA expression as well as gene expression when RNA biomarkers turn genes on. We discussed the real-cell biology of miRNA biomarkers near the heart to allow us to further understand the concentration of miRNAs that circulate in the blood.

Dr. Charles Searles is a cardiologist specialist working with biomarkers regarding CAD and an associate professor of medicine at Emory Healthcare Hospital. He provided us with an additional miRNA as a control for our proof of concept experimentation. He also offered to test our padlocks using Digital PCR.

Dr. Ugochukwu Egolum is a cardiologist specialist in Gainesville, Georgia, and is affiliated with multiple hospitals in Northeast Georgia. He helped our research on miRNAs as he discussed the purposes of each miRNA we had researched, signifying which miRNAs were consistent with diagnosing CAD.

CAD Researchers

Dr. Omar Kashlan is a cardiologist working at Wellstar Hospital in downtown Atlanta. He discussed the benefits of early diagnosis of CAD, which consist of alternative manifestations such as heart attacks. This early screening will be beneficial for the patients and can help doctors ensure accurate results as elevated miRNA levels could be helpful in detection in case abnormal levels of calcium are not present.

Dr. Wissam Jaber is a cardiologist, who specializes in Coronary Artery Disease at Emory Healthcare Hospital. He explained the current trends of the diagnosis of CAD which are for less invasive methods and identifying biomarkers that align with the disease. Our project aligns with both trends, leading us to question current methods of diagnosis.

Dr. Alison Ward is a cardiac surgeon at Grady Hospital. She explained that high sensitivity troponin levels were a current trend in cardiac diagnostics. She also explained how all patients are different as female patients have less reliable exams such as epigastric pain which makes the diagnostic test results less precise. We took this into account to ensure that all patients have accurate results even from their different symptoms, allowing for a more diverse administration of our test.

Human Practices

The Lambert High School culinary team helped us find diverse heart-healthy recipes for our cookbook, test them, and provide accurate and desired nutritional information about each recipe.

The Georgia Office of Cardiac Health helped us learn about data regulations in our state of Georgia, verified that coronary artery disease was the most prevalent condition in the state, and collaborated to create an education pamphlet that would be displayed in Georgia offices and clinics to answer common misconceptions.

Piper Allsup is a student at Lambert High School who greatly contributed to the development of our cookbook. She helped us research many recipes and ultimately worked to make our cookbook filled with lots of information about recipes found around the world.

Inclusivity

At Lambert High School, Karen Surio RN, the school nurse; Ann Berry, speech-language pathologist; and Jennifer Ro, county-level speech pathologist, work with special needs students. They gave suggestions of words that they thought would be beneficial to have on the communication board. In addition, we received feedback and approval of our final product. 

Dr. Charles Searles from Emory Healthcare Hospital discussed the feasibility of building a database. He helped us design CADmiR and gave us advice on what to include to cover a user's desired information about miRNAs.

Model

Dr. Mark Styczynski explained what equations would or wouldn’t work for rolling circle amplification (RCA) and rolling circle transcription (RCT) models and where to theoretically start if there is a lack of probable results from wet lab.

Wetlab

Padlock Probes

Dr. Mats Nilsson helped us determine the feasibility of padlocks for miRNA detection and guided us in the development of our first padlock probe.

Megan McSweeney from Styczinski Lab at Georgia Institute of Technology helped us troubleshoot linear probes and any errors with aptamers.

Proposed Implementation

Dr. Christian Delles is a professor and head of school at the University of Glasgow who specializes in cardiovascular and metabolic health. He helped us conduct the meta-study by guiding us through on what information needed to be included in the data set for a successful conclusion on microRNA levels for healthy patients and patients with CAD. 

RCT

Dr. Mats Nilsson introduced us to rolling circle transcription (RCT) and assisted us in learning more about the topic and how it works.

Dr. Erik T. Kool from Kool Lab in Stanford taught us how to troubleshoot rolling circle transcription and helped us through the troubleshoot process of RCT.

Alexandra Patterson from the Georgia Institute of Technology sent us the three-way junction broccoli and guided RCT with broccoli aptamer use.

RCA

Dr. Mark Styczynski from Styczinski Lab at Georgia Institute of Technology helped us troubleshoot rolling circle amplification controls.

Dr. Jeremy Koob is a graduate student from Harvard University who worked with SHERLOCK, however he confirmed that RCA worked better and he helped us justify why it led to better and more accurate results.

Entrepreneurship

Dr. Ahmed Alawad was a former business development manager for the Middle Eastern and North African (MENA) region for Johnson and Johnson and a startup founder in Atlanta, Georgia. After examining our business plan, he discussed product credibility, seed funding options, and financial allocation.

Mr. John Nahas is an investor and startup founder based in Los Angles, California, who founded U.C Berkeley's first venture capital club and graduated from Berkeley HAAS Business School. He critiqued our industry analysis and helped us improve consumer perception, sales, financing strategy, and revenue.

Dr. Viju Kulkarni is a retired economics professor from San Diego State University. Dr. Kulkarni helped us learn about what goes into making a successful business plan and how to conduct market research.

Hardware

Dr. Saad Bhamla - Bhamla Lab, Georgia Institute of Technology: Dr. Bhamla held monthly meetings to guide our Micro-Q team to refine their designs and provided constructive feedback on our work in 2022.

Rajas Poorna - PhD Student, Georgia Institute of Technology: Mr. Poorna led troubleshooting sessions that helped us solve issues such as leaky fluorescence in Micro-Q and system miniaturization.

Other iGEM Teams

Ribotox - City_of_London_UK, 2021: Ribotox provided evidence that microRNA biosensing was feasible in a high school lab.

Viral Spiral - TAS_Taipei, 2020: Viral Spiral provided a foundation for Lambert iGEM's rolling circle amplification and transcription biosensing approaches for RNA targets.