title: Mouse genome engineering via CRISPR-Cas9 for study of immune function.
abstract: Clustered regularly interspaced palindromic repeats (CRISPR)-associated (Cas9) technology has proven a formidable addition to our armory of approaches for genomic editing. Derived from pathways in archaea and bacteria that mediate the resistance to exogenous genomic material, the CRISPR-Cas9 system utilizes a short single guide RNA (sgRNA) to direct the endonuclease Cas9 to virtually anywhere in the genome. Upon targeting, Cas9 generates DNA double-strand breaks (DSBs) and facilitates the repair or insertion of mutations, insertion of recombinase recognition sites, or large DNA elements. Here, we discuss the practical advantages of the CRISPR-Cas9 system over conventional and other nuclease-based targeting technologies and provide suggestions for the use of this technology to address immunological questions.
authors.affiliation: Department of Immunology, St. Jude Children's Hospital, Memphis, TN 38103, USA. Electronic address: stephane.pelletier@stjude.org.
authors.forename: Stephane
authors.initials: S
authors.lastname: Pelletier
authors.affiliation: Department of Immunology, St. Jude Children's Hospital, Memphis, TN 38103, USA.
authors.forename: Sebastien
authors.initials: S
authors.lastname: Gingras
authors.affiliation: Department of Immunology, St. Jude Children's Hospital, Memphis, TN 38103, USA. Electronic address: douglas.green@stjude.org.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Green
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004260:DNA Repair; D004720:Endonucleases / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D016678:Genome / Q000235:genetics; D006801:Humans; D007109:Immunity / Q000235:genetics*; D051379:Mice; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D004720:Endonucleases