CLICK: one-step generation of conditional knockout mice.

ABSTRACT: BACKGROUND:CRISPR/Cas9 enables the targeting of genes in zygotes; however, efficient approaches to create loxP-flanked (floxed) alleles remain elusive. RESULTS:Here, we show that the electroporation of Cas9, two gRNAs, and long single-stranded DNA (lssDNA) into zygotes, termed CLICK (CRISPR with lssDNA inducing conditional knockout alleles), enables the quick generation of floxed alleles in mice and rats. CONCLUSIONS:The high efficiency of CLICK provides homozygous knock-ins in oocytes carrying tissue-specific Cre, which allows the one-step generation of conditional knockouts in founder (F0) mice.

SUBMITTER: Miyasaka Y 

PROVIDER: S-EPMC5930688 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature