A Co-CRISPR Strategy for Efficient Genome Editing in Caenorhabditis Elegans

Overview

Journal Genetics

Publisher Oxford University Press

Specialty Genetics

Date 2014 Jun 1

PMID 24879462

Citations 178

Authors

Heesun Kim

Takao Ishidate

Krishna S Ghanta

Meetu Seth

Darryl Conte Jr

Masaki Shirayama

Craig C Mello

Affiliations

Soon will be listed here.

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.

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