A universal method for generating knockout mice in multiple genetic backgrounds using zygote electroporation.

Autor:	Tamari T; Model Generation & Breeding Service, The Jackson Laboratory Japan, Inc., 955 Kamibayashi, Ishioka, Ibaraki 315-0138, Japan.; Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.; Doctoral Program in Biomedical Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan., Ikeda Y; Model Generation & Breeding Service, The Jackson Laboratory Japan, Inc., 955 Kamibayashi, Ishioka, Ibaraki 315-0138, Japan.; Laboratory Animal Resource Center in Trans-Border Medical Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan., Morimoto K; Doctoral Program in Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.; Research Fellow of the Japan Society for the Promotion of Science, Kojimachi Business Center Building, 5-3-1 Kojimachi, Chiyoda-ku, Tokyo 102-0083, Japan., Kobayashi K; Model Generation & Breeding Service, The Jackson Laboratory Japan, Inc., 955 Kamibayashi, Ishioka, Ibaraki 315-0138, Japan., Mizuno-Iijima S; Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan., Ayabe S; Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan., Kuno A; Department of Anatomy and Embryology, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan., Mizuno S; Laboratory Animal Resource Center in Trans-Border Medical Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan., Yoshiki A; Experimental Animal Division, RIKEN BioResource Research Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
Jazyk:	angličtina
Zdroj:	Biology open [Biol Open] 2023 Sep 15; Vol. 12 (9). Date of Electronic Publication: 2023 Aug 25.
DOI:	10.1242/bio.059970
Abstrakt:	Genetically engineered mouse models are essential tools for understanding mammalian gene functions and disease pathogenesis. Genome editing allows the generation of these models in multiple inbred strains of mice without backcrossing. Zygote electroporation dramatically removed the barrier for introducing the CRISPR-Cas9 complex in terms of cost and labour. Here, we demonstrate that the generalised zygote electroporation method is also effective for generating knockout mice in multiple inbred strains. By combining in vitro fertilisation and electroporation, we obtained founders for knockout alleles in eight common inbred strains. Long-read sequencing analysis detected not only intended mutant alleles but also differences in read frequency of intended and unintended alleles among strains. Successful germline transmission of knockout alleles demonstrated that our approach can establish mutant mice targeting the same locus in multiple inbred strains for phenotyping analysis, contributing to reverse genetics and human disease research. Competing Interests: Competing interests T.T., K.K. and Y.I. are employed by The Jackson Laboratory Japan, Inc., which provided the mice described in the paper. The Jackson Laboratory Japan, Inc. is not a publicly traded company and the authors do not own any shares/equity in the company. (© 2023. Published by The Company of Biologists Ltd.)
Databáze:	MEDLINE
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