title: Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing.
abstract: The CRISPR-Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR-Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double-strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR-Cas9 system.
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Wenning
authors.initials: W
authors.lastname: Qin
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Kutny
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Richard S
authors.initials: RS
authors.lastname: Maser
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Dion
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Lamont
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Yingfan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.
authors.forename: Greggory A
authors.initials: GA
authors.lastname: Perry
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine.|State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Haoyi
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D023281:Genomics / Q000379:methods*; D060005:Genotyping Techniques; D051379:Mice; D008845:Microinjections; D023421:Models, Animal; D009841:Oligonucleotides; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D015053:Zygote
chemical_list: D009841:Oligonucleotides; D012333:RNA, Messenger
keywords: CRISPR; genome editing; knock-in; knockout; mouse model