A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans
Document Type
Journal ArticlePublication Date
2014-08-01Keywords
Animals; Base Sequence; CRISPR-Associated Proteins; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Clustered Regularly Interspaced Short Palindromic Repeats; Deoxyribonucleases; Genetic Markers; *Genome, Helminth; Homologous Recombination; Molecular Sequence Data; Mutagenesis, Insertional; Plasmids; Sequence Analysis, DNACRISPR-Cas9 system
Co-CRISPR
CRISPR-Cas9-mediated HR
CRISPR-Cas9-induced indels
blasticidin selection
Computational Biology
Genomics
Molecular Biology
Molecular Genetics
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Show full item recordAbstract
Genome editing based on CRISPR (clustered regularly interspaced short palindromic repeats)-associated nuclease (Cas9) has been successfully applied in dozens of diverse plant and animal species, including the nematode Caenorhabditis elegans. The rapid life cycle and easy access to the ovary by micro-injection make C. elegans an ideal organism both for applying CRISPR-Cas9 genome editing technology and for optimizing genome-editing protocols. Here we report efficient and straightforward CRISPR-Cas9 genome-editing methods for C. elegans, including a Co-CRISPR strategy that facilitates detection of genome-editing events. We describe methods for detecting homologous recombination (HR) events, including direct screening methods as well as new selection/counterselection strategies. Our findings reveal a surprisingly high frequency of HR-mediated gene conversion, making it possible to rapidly and precisely edit the C. elegans genome both with and without the use of co-inserted marker genes.Source
Genetics. 2014 Aug;197(4):1069-80. doi: 10.1534/genetics.114.166389. Epub 2014 May 30. Link to article on publisher's siteDOI
10.1534/genetics.114.166389Permanent Link to this Item
http://hdl.handle.net/20.500.14038/33369PubMed ID
24879462Related Resources
Link to Article in PubMedRights
Copyright © 2014 by the Genetics Society of America. Available freely online through the author-supported open access option.
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