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

Chen, Changchun; Fenk, Lorenz A.; de Bono, Mario

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

Chen C, Fenk LA, de Bono M. 2013. Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. Nucleic Acids Research. 41(20), e193.

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DOI

10.1093/nar/gkt805

Journal Article | Published | English

Author

Chen, Changchun; Fenk, Lorenz A.; de Bono, Mario^ISTA

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.

Publishing Year

2013

Date Published

2013-11-01

Journal Title

Nucleic Acids Research

Publisher

Oxford University Press

Acknowledgement

The authors thank the Caenorhabditis Genetics Center for strains, Erik Jorgensen and Jason Chin for plasmids, Sebastian Greiss and Inja Radman for valuable suggestions and Shigekazu Oda for comments on the article. The Ce Cas9 expression plasmid and ‘empty’ versions of the two sgRNA expression plasmids described in the article are available at Addgene.org.

Volume

Issue

Article Number

e193

ISSN

0305-1048

eISSN

1362-4962

IST-REx-ID

6130

Cite this

Chen C, Fenk LA, de Bono M. Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. Nucleic Acids Research. 2013;41(20). doi:10.1093/nar/gkt805

Chen, C., Fenk, L. A., & de Bono, M. (2013). Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. Nucleic Acids Research. Oxford University Press. https://doi.org/10.1093/nar/gkt805

Chen, Changchun, Lorenz A. Fenk, and Mario de Bono. “Efficient Genome Editing in Caenorhabditis Elegans by CRISPR-Targeted Homologous Recombination.” Nucleic Acids Research. Oxford University Press, 2013. https://doi.org/10.1093/nar/gkt805.

C. Chen, L. A. Fenk, and M. de Bono, “Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination,” Nucleic Acids Research, vol. 41, no. 20. Oxford University Press, 2013.

Chen C, Fenk LA, de Bono M. 2013. Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. Nucleic Acids Research. 41(20), e193.

Chen, Changchun, et al. “Efficient Genome Editing in Caenorhabditis Elegans by CRISPR-Targeted Homologous Recombination.” Nucleic Acids Research, vol. 41, no. 20, e193, Oxford University Press, 2013, doi:10.1093/nar/gkt805.

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