| Literature DB >> 26068112 |
Nicolas Wyvekens1,2, Ved V Topkar1,2, Cyd Khayter1,2, J Keith Joung1,2,3, Shengdar Q Tsai1,2,3.
Abstract
Monomeric clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated 9 (Cas9) nucleases have been widely adopted for simple and robust targeted genome editing but also have the potential to induce high-frequency off-target mutations. In principle, two orthogonal strategies for reducing off-target cleavage, truncated guide RNAs (tru-gRNAs) and dimerization-dependent RNA-guided FokI-dCas9 nucleases (RFNs), could be combined as tru-RFNs to further improve genome editing specificity. Here we identify a robust tru-RFN architecture that shows high activity in human cancer cell lines and embryonic stem cells. Additionally, we demonstrate that tru-gRNAs reduce the undesirable mutagenic effects of monomeric FokI-dCas9. Tru-RFNs combine the advantages of two orthogonal strategies for improving the specificity of CRISPR-Cas nucleases and therefore provide a highly specific platform for performing genome editing.Entities:
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Year: 2015 PMID： 26068112 PMCID： PMC4509490 DOI： 10.1089/hum.2015.084
Source DB: PubMed Journal: Hum Gene Ther ISSN： 1043-0342 Impact factor: 5.695