UMMS Affiliation
Program in Molecular Medicine; RNA Therapeutics Institute
Publication Date
2014-08-01
Document Type
Article
Disciplines
Computational Biology | Genomics | Molecular Biology | Molecular Genetics
Abstract
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.
Keywords
CRISPR-Cas9 system, Co-CRISPR, CRISPR-Cas9-mediated HR, CRISPR-Cas9-induced indels, blasticidin selection
Rights and Permissions
Copyright © 2014 by the Genetics Society of America. Available freely online through the author-supported open access option.
DOI of Published Version
10.1534/genetics.114.166389
Source
Genetics. 2014 Aug;197(4):1069-80. doi: 10.1534/genetics.114.166389. Epub 2014 May 30. Link to article on publisher's site
Journal/Book/Conference Title
Genetics
Related Resources
PubMed ID
24879462
Repository Citation
Kim H, Ishidate T, Ghanta KS, Seth M, Conte D, Shirayama M, Mello CC. (2014). A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans. Morningside Graduate School of Biomedical Sciences Student Publications. https://doi.org/10.1534/genetics.114.166389. Retrieved from https://escholarship.umassmed.edu/gsbs_sp/1893
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