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|
| Type |
Poster Presentation |
| Area |
Life Chemistry |
| Room No. |
Event Hall |
| Time |
4월 20일 (금요일) 11:00~12:30 |
| Code |
BIO.P-263 |
| Subject |
Incorporation of bridged nucleic acid into CRISPR RNA enhances target specificity of Cas9 endonuclease |
| Authors |
Jinho Park, Keewon Sung, Christopher Cromwell1, Amanda Krysler1, Juan Jovel2, Basil Hubbard1,*, Seong Keun Kim*
Department of Chemistry, Seoul National University, Korea
1Department of Pharmacology, University of Alberta, Canada
2The Applied Genomics Core, Office of Research, Canada
|
| Abstract |
Application of the CRISPR/Cas9 technology to functional genetics and human therapeutics is hindered by its unintended off-target DNA cleavage effect. Here, we show that the incorporation of next-generation bridged nucleic acids (2’, 4’ BNANC[N-Me]) into CRISPR-RNAs (crRNAs) significantly reduces off-target DNA cleavage by Cas9 in vitro and in cells. In addition, using single-molecule FRET technique, we show that the incorporation of BNA can affect dynamical aspects as well, including the crRNA-related conformational change between two distinct states with ‘open’ and ‘zipped’ structures. These results raise the prospect of highly target-specific gene-editing techniques using the CRISPR/Cas9 system and may shed light on new applications of synthetic nucleic acids. |
| E-mail |
jpark93@snu.ac.kr |
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121st General Meeting of the KCS