RRC ID 45770
著者 Kim H, Ishidate T, Ghanta KS, Seth M, Conte D Jr, Shirayama M, Mello CC.
タイトル A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans.
ジャーナル 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.
巻・号 197(4)
ページ 1069-80
公開日 2014-8-1
DOI 10.1534/genetics.114.166389
PII genetics.114.166389
PMID 24879462
PMC PMC4125384
MeSH Animals Base Sequence CRISPR-Associated Proteins / genetics* Caenorhabditis elegans / genetics* Caenorhabditis elegans Proteins / genetics* Clustered Regularly Interspaced Short Palindromic Repeats / genetics* Deoxyribonucleases / genetics* Genetic Markers Genome, Helminth* Homologous Recombination / genetics Molecular Sequence Data Mutagenesis, Insertional Plasmids / genetics Sequence Analysis, DNA
IF 4.015
引用数 120
WOS 分野 GENETICS & HEREDITY
リソース情報
線虫 tm1396