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Efficient CRISPR/Cas9-Mediated Genome Editing in Mice by Zygote Electroporation of Nuclease. Genetics 2015 Jun;200(2):423-30

Date

03/31/2015

Pubmed ID

25819794

Pubmed Central ID

PMC4492369

DOI

10.1534/genetics.115.176594

Scopus ID

2-s2.0-84931385060   91 Citations

Abstract

The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system is an adaptive immune system in bacteria and archaea that has recently been exploited for genome engineering. Mutant mice can be generated in one step through direct delivery of the CRISPR/Cas9 components into a mouse zygote. Although the technology is robust, delivery remains a bottleneck, as it involves manual injection of the components into the pronuclei or the cytoplasm of mouse zygotes, which is technically demanding and inherently low throughput. To overcome this limitation, we employed electroporation as a means to deliver the CRISPR/Cas9 components, including Cas9 messenger RNA, single-guide RNA, and donor oligonucleotide, into mouse zygotes and recovered live mice with targeted nonhomologous end joining and homology-directed repair mutations with high efficiency. Our results demonstrate that mice carrying CRISPR/Cas9-mediated targeted mutations can be obtained with high efficiency by zygote electroporation.

Author List

Qin W, Dion SL, Kutny PM, Zhang Y, Cheng AW, Jillette NL, Malhotra A, Geurts AM, Chen YG, Wang H

Authors

Yi-Guang Chen PhD Associate Professor in the Pediatrics department at Medical College of Wisconsin
Aron Geurts PhD Associate Professor in the Physiology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Base Sequence
CRISPR-Cas Systems
Electroporation
Endonucleases
Female
Gene Targeting
Genetic Loci
Genome
Genomics
INDEL Mutation
Mice
Molecular Sequence Data
RNA Editing
RNA, Guide
RNA, Messenger
Sequence Alignment
Zygote
jenkins-FCD Prod-398 336d56a365602aa89dcc112f077233607d6a5abc