Generating Mouse Models Using Zygote Electroporation of Nucleases (ZEN) Technology with High Efficiency and Throughput.

Document Type

Article

Publication Date

1-1-2017

Publication Title

Methods in molecular biology (Clifton, N.J.)

Keywords

JMG, Animals, CRISPR-Cas Systems, Electroporation, Endonucleases, Mice, Mice, Transgenic, Models, Animal, Zygote

JAX Source

Methods Mol Biol . 2017:1605:219-230

Volume

1605

First Page

219

Last Page

230

ISSN

1940-6029

PMID

28456968

DOI

10.1007/978-1-4939-6988-3_15

Abstract

Mouse models with genetic modifications are widely used in biology and biomedical research. Although the application of CRISPR-Cas9 system greatly accelerated the process of generating genetically modified mice, the delivery method depending on manual injection of the components into the embryos remains a bottleneck, as it is laborious, low throughput, and technically demanding. To overcome this limitation, we invented and optimized the ZEN (Zygote electroporation of nucleases) technology to deliver CRISPR-Cas9 reagents via electroporation. Using ZEN, we were able to generate genetically modified mouse models with high efficiency and throughput. Here, we describe the protocol in great detail.

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