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Curr Protoc Mouse Biol. 2016 Mar 1;6(1):39-66. doi: 10.1002/9780470942390.mo150178.

Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing.

Author information

1
The Jackson Laboratory, Bar Harbor, Maine.
2
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

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.

KEYWORDS:

CRISPR; genome editing; knock-in; knockout; mouse model

PMID:
26928663
PMCID:
PMC4848752
[Available on 2017-03-01]
DOI:
10.1002/9780470942390.mo150178
[Indexed for MEDLINE]
Free PMC Article

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