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Nucleic Acids Res. 2013 Nov;41(20):e193. doi: 10.1093/nar/gkt805. Epub 2013 Sep 5.

Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination.

Author information

1
Division of Cell Biology, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.

Abstract

Cas9 is an RNA-guided double-stranded DNA nuclease that participates in clustered regularly interspaced short palindromic repeats (CRISPR)-mediated adaptive immunity in prokaryotes. CRISPR-Cas9 has recently been used to generate insertion and deletion mutations in Caenorhabditis elegans, but not to create tailored changes (knock-ins). We show that the CRISPR-CRISPR-associated (Cas) system can be adapted for efficient and precise editing of the C. elegans genome. The targeted double-strand breaks generated by CRISPR are substrates for transgene-instructed gene conversion. This allows customized changes in the C. elegans genome by homologous recombination: sequences contained in the repair template (the transgene) are copied by gene conversion into the genome. The possibility to edit the C. elegans genome at selected locations will facilitate the systematic study of gene function in this widely used model organism.

PMID:
24013562
PMCID:
PMC3814388
DOI:
10.1093/nar/gkt805
[Indexed for MEDLINE]
Free PMC Article

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