G-quadruplex motif at the 3' end of sgRNAs improves CRISPR-Cas9 based genome editing efficiency
| Title | G-quadruplex motif at the 3' end of sgRNAs improves CRISPR-Cas9 based genome editing efficiency |
| Publication Type | Journal Article |
| Year of Publication | 2018 |
| Authors | Nahar, S, Sehgal, P, Azhar, M, Rai, M, Singh, A, Sivasubbu, S, Chakraborty, D, Maiti, S |
| Journal | Chemical Communications |
| Volume | 54 |
| Issue | 19 |
| Pagination | 2377-2380 |
| Date Published | MAR |
| ISSN | 1359-7345 |
| Abstract | Originating as a component of prokaryotic adaptive immunity, the type II CRISPR/Cas9 system has been repurposed for targeted genome editing in various organisms. Although Cas9 can bind and cleave DNA efficiently under in vitro conditions, its activity inside a cell can vary dramatically between targets owing to differences between genomic loci and availability of enough Cas9/sgRNA (single guide RNA) complex molecules for cleavage. Most methods to improve CRISPR/Cas9 activity have so far relied on Cas9 protein engineering or base modifications in the sgRNA sequence. Here we demonstrate that a structure based rational design of sgRNAs can enhance the efficiency of Cas9 cleavage in vivo. By appending a naturally forming RNA G-quadruplex motif to the 3′ end of sgRNAs we can improve its stability and target cleavage efficiency in zebrafish embryos without inducing off-target activity, thereby underscoring its value in the design of better and optimized genome editing triggers. |
| DOI | 10.1039/C7CC08893K |
| Type of Journal (Indian or Foreign) | Foreign |
| Impact Factor (IF) | 6.319 |
Divison category:
Biochemical Sciences
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