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| | <br/>DESCRIPTION</div></div> | | <br/>DESCRIPTION</div></div> |
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| − | <p align="justify">Since we had a great summer last year working on a project about the new application of the genome editing technique TALEN. This year, in our brainstorming process, we decided to stick to this strategy and to discover the brand-new application of genome editing techniques on other purposes of usage. </p> | + | <p align="justify">MicroRNAs, serve as critical gene expression regulators at the transcriptional and post-transcriptional levels, have also been found as important blood-based biomarkers for early detection of cancers. However, their current in vitro detection methods are relatively complex, costly and low sensitive. Our project attempts to establish a novel in vitro microRNA detection system which is rapid, efficient, sensitive and specific. In this system, CRISPR-Cas9 technique is modified to integrate with split-luciferase or split-HRP reporting systems. The advanced rolling circle amplification technology and cell-free expression system are also involved and optimized. This system may ideally be compatible for the detection of various series of small non-coding RNAs. To our knowledge, we are the first to use the CRISPR-Cas9 system as a small non-coding RNA monitor in vitro. Its establishment and further development might provide a new approach for rapid and low-cost cancer screening, virus detection and curative efficacy assessment. </p></div> |
| − | <p align="justify">In our literature research, we found out that Small non-coding RNAs (sncRNAs, invluding microRNAs, piRNAs etc.), as a family of non-coding RNAs with the length no larger than 200nt, has now been reported relevant with the tumorigenesis, development and metastasis of several different cancers. Some also reported sncRNAs as promising bio-markers for the early detection of several specific cancers. However, the rapid and low-cost detection of sncRNAs remains problematic, thus limits the further implementation of sncRNAs in the early detection of cancers. Meanwhile, the lack of valid tools for the real-time and in vivo detection of sncRNAs in living cells also limits the further development of cancer research.</p>
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| − | <p align="justify">In our project this year, we intend to use the rapid-developing CRISPR/Cas9 technique in combination with other genome-editing tools such as Zinc Finger Protein to develop a new method for the detection of sncRNAs by using the nucleic acid binding ability of such tools. The split-GFP, split-luciferase, and solit-HRP systems will be the candidate manners for generation of reporting signal in our study. We have now conducted in vitro Cas9 cutting experiments for the primary test of our design, and to select the sncRNA for our prototype. Our primary results turned out positively. Also, we constructed and compared two different split-GFP systems, to select the best one for our further study. The split-HRP and split-Luciferase system are also being tested.</p>
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| − | <p align="justify">We believe that our system can be a brand new application of CRISPR/Cas9 system and can provide a rapid, low-cost platform for the detection of sncRNAs. We also hope to see the further extension of this project on the early diagnosis of cancers.
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