Article
CLICK: one-step generation of conditional knockout mice.
BMC genomics - 2 May 2018
Miyasaka Yoshiki, Uno Yoshihiro, Yoshimi Kazuto, Kunihiro Yayoi, Yoshimura Takuji, Tanaka Tomohiro, Ishikubo Harumi, Hiraoka Yuichi, Takemoto Norihiko, Tanaka Takao, Ooguchi Yoshihiro, Skehel Paul, Aida Tomomi, Takeda Junji, Mashimo Tomoji
Abstract excerpt
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....
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