Multiplex Genome Engineering Using CRISPR/Cas Systems
Top Cited Papers
Open Access
- 15 February 2013
- journal article
- other
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 339 (6121), 819-823
- https://doi.org/10.1126/science.1231143
Abstract
Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.This publication has 32 references indexed in Scilit:
- Cas9–crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteriaProceedings of the National Academy of Sciences of the United States of America, 2012
- FLASH assembly of TALENs for high-throughput genome editingNature Biotechnology, 2012
- Tracking genome engineering outcome at individual DNA breakpointsNature Methods, 2011
- Evolution and classification of the CRISPR–Cas systemsNature Reviews Microbiology, 2011
- CRISPR RNA maturation by trans-encoded small RNA and host factor RNase IIINature, 2011
- Efficient construction of sequence-specific TAL effectors for modulating mammalian transcriptionNature Biotechnology, 2011
- Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA)Nature Methods, 2010
- Small RNAs as Guardians of the GenomeCell, 2009
- An improved zinc-finger nuclease architecture for highly specific genome editingNature Biotechnology, 2007
- Mechanisms of gene silencing by double-stranded RNANature, 2004