Heritable Gene Knockout in Caenorhabditis elegans by Direct Injection of Cas9–sgRNA Ribonucleoproteins

We present a novel method of targeted gene disruption that involves direct injection of recombinant Cas9 protein complexed with guide RNA into the gonad of the nematode Caenorhabditis elegans. Biallelic mutants were recovered among the F1 progeny, demonstrating the high efficiency of this method.

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Bibliographic Details
Published inGenetics (Austin) Vol. 195; no. 3; pp. 1177 - 1180
Main Authors Cho, Seung Woo, Lee, Jihyun, Carroll, Dana, Kim, Jin-Soo, Lee, Junho
Format Journal Article
LanguageEnglish
Published United States Genetics Society of America 01.11.2013
Subjects
Animals
Base Sequence
Caenorhabditis elegans
Caenorhabditis elegans - genetics
CRISPR-Cas Systems
DNA, Helminth - genetics
Gene Knockout Techniques
Gene Targeting
Genes
Genes, Helminth
Genetics
Injection
Molecular Sequence Data
Nematoda
Nematodes
Ribonucleic acid
RNA
Caenorhabditis elegans
genome editing
Cas9–sgRNA ribonucleoprotein
NHEJ
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Summary:We present a novel method of targeted gene disruption that involves direct injection of recombinant Cas9 protein complexed with guide RNA into the gonad of the nematode Caenorhabditis elegans. Biallelic mutants were recovered among the F1 progeny, demonstrating the high efficiency of this method.
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Supporting information is available online at http://www.genetics.org/lookup/suppl/doi:10.1534/genetics.113.155853/-/DC1.
These authors contributed equally to this work.
ISSN:1943-2631
0016-6731
1943-2631
DOI:10.1534/genetics.113.155853