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Delivery of Cas9 Protein into Mouse Zygotes through a Series of Electroporation Dramatically Increases the Efficiency of Model Creation

Author:
Wang, Wenbo, Kutny, Peter M., Byers, Shannon L., Longstaff, Charles J., DaCosta, Michael J., Pang, Changhong, Zhang, Yingfan, Taft, Robert A., Buaas, Frank W., Wang, Haoyi
Source:
Journal of genetics and genomics 2016 v.43 no.5 pp. 319-327
ISSN:
1673-8527
Subject:
animal models, electroporation, loci, mice, nucleases, transgenic animals, zygote
Abstract:
Previously we established Zygote Electroporation of Nucleases (ZEN) technology as an efficient and high-throughput way to generate genetically modified mouse models. However, there were significant variations of the targeting efficiency among different genomic loci using our previously published protocol. In this study, we improved the ZEN technology by delivering Cas9 protein into mouse zygotes through a series of electroporation. Using this approach, we were able to introduce precise nucleotide substitutions, large segment deletion and short segment insertion into targeted loci with high efficiency.
Agid:
5306335