RRC ID 50502
Author Miyasaka Y, Uno Y, Yoshimi K, Kunihiro Y, Yoshimura T, Tanaka T, Ishikubo H, Hiraoka Y, Takemoto N, Tanaka T, Ooguchi Y, Skehel P, Aida T, Takeda J, Mashimo T.
Title CLICK: one-step generation of conditional knockout mice.
Journal BMC Genomics
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.
Volume 19(1)
Pages 318
Published 2018-5-2
DOI 10.1186/s12864-018-4713-y
PII 10.1186/s12864-018-4713-y
PMID 29720086
PMC PMC5930688
MeSH Alleles Animals Base Sequence CRISPR-Cas Systems / genetics Genetic Engineering / methods* Injections Mice Mice, Knockout Zygote / metabolism
IF 3.594
Mice RBRC01828