Overview
Since the beginning of the project, it has always been our theme to make useful contributions to the future iGEM team. We believe that the experience accumulated by each iGEM team is very meaningful for the future iGEMer. The last iGEMer has transcribed a lot of content in their wiki, and we have benefited a lot, so we decided to do our best to help the next iGEMer. Our main contribution to the future iGEM team is the design of HCR system and CRISPR system. In addition, we also list other elements below that we think may be useful to anyone interested.
Probe Design for HCR and NUPACK User Guide
Selecting or reasonably designing HCR probes to connect CRISPR functions together is a key and arduous step. In the past few months, we have worked together to design and test many different probe designs, and we are finally happy to share our results with the iGEM community.
In our HCR reaction, we designed three probes H1, H2, and H3 for experiments. Since the base structure, stem-loop length, base mismatch, etc. of the probe will affect the amplification effect of the HCR reaction, in the modeling work, we design probes with different base sequences, stem-loop length, and base mismatch, and simulate them with NUPACK to analyze the free energy of the product and the equilibrium concentration of the system, and then evaluate the amplification performance of the HCR reaction.
To help other teams that need to optimize the probe, we have produced a NUPACK user manual.
Laboratory manual on HCR and CRISPR
We have prepared an experimental manual on HCR and CRISPR. In fact, it includes not only the basic experimental operations, but also some experimental improvement schemes used to optimize the results. We hope that this manual can help teams who want to adopt this technology.
Paper Based Chip Model
We use dry HCR reagents to react on paper materials and realize the biological system of HCR+CRISPR on paper chips. We shared the materials and processing steps of paper chips to enable other teams to reproduce our paper chips.
Instant Detection Operating Platform Based On Smart Phone
Diagnosis is complicated by the presence of chronic musculoskeletal pain, because of a mixture of neuropathic and nociceptive pathogens, or inflammatory signs associated with chronic regional pain syndrome (CRPS). Because there is a strong correlation between treatment and nociceptive pain or neuropathic pain, this project has developed a miniaturized, integrated, automated, interactive device that detects relevant biomarker miRNAs. The device uses the App to quantitatively analyze the image to obtain fluorescence intensity information, and analyzes the corresponding concentration of miRNA, providing objective detection data to assist doctors in classifying and diagnosing diseases, which is convenient for further treatment.
3D Modeling of Hardware and Prompt for Documentation
We have carried out 3D modeling for the hardware part and written the description documents of relevant parts. Both the incubation device structure and the fluorescence detection structure can provide design inspiration for other teams with similar heating or fluorescence detection needs. Our paper chip carrier could also serve other teams with similar needs by simply changing the length, width, and height of the structure to fit their devices. Because the structure can be manufactured by 3D printing, as long as it can meet the requirements of experimental conditions, we can use recycled plastic as raw materials for manufacturing. Finally, the mass production of the structure can be realized by making molds.
Classification Model
Our study obtained a miRNA set with a high correlation with pain (miRNA in the 14 above) and 119 miRNAs with a high concentration in blood, which can provide a suitable range of marker selection for similar experimental studies and provide a reference for other miRNA-related detection technologies. For many miRNA detection work to avoid a lot of complicated screening stages. In addition, based on our screening process, the model, and the corresponding literature data, we hope that the next miRNAs & pain research projects can be further on our work with a table of miRNAs & pain, then we may be able to explain the mechanism of action of pain from the perspective of genetic, and explore the underlying etiology, Even with the help of the control table to develop miRNA analgesic drugs.