Back to Search
Start Over
Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants.
- Source :
-
Science (New York, N.Y.) [Science] 2020 Apr 17; Vol. 368 (6488), pp. 290-296. Date of Electronic Publication: 2020 Mar 26. - Publication Year :
- 2020
-
Abstract
- Manipulation of DNA by CRISPR-Cas enzymes requires the recognition of a protospacer-adjacent motif (PAM), limiting target site recognition to a subset of sequences. To remove this constraint, we engineered variants of Streptococcus pyogenes Cas9 (SpCas9) to eliminate the NGG PAM requirement. We developed a variant named SpG that is capable of targeting an expanded set of NGN PAMs, and we further optimized this enzyme to develop a near-PAMless SpCas9 variant named SpRY (NRN and to a lesser extent NYN PAMs). SpRY nuclease and base-editor variants can target almost all PAMs, exhibiting robust activities on a wide range of sites with NRN PAMs in human cells and lower but substantial activity on those with NYN PAMs. Using SpG and SpRY, we generated previously inaccessible disease-relevant genetic variants, supporting the utility of high-resolution targeting across genome editing applications.<br /> (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
Details
- Language :
- English
- ISSN :
- 1095-9203
- Volume :
- 368
- Issue :
- 6488
- Database :
- MEDLINE
- Journal :
- Science (New York, N.Y.)
- Publication Type :
- Academic Journal
- Accession number :
- 32217751
- Full Text :
- https://doi.org/10.1126/science.aba8853