CLICK: One-step generation of conditional knockout mice

Yoshiki Miyasaka, Yoshihiro Uno, Kazuto Yoshimi, Yayoi Kunihiro, Takuji Yoshimura, Tomohiro Tanaka, Harumi Ishikubo, Yuichi Hiraoka, Norihiko Takemoto, Takao Tanaka, Yoshihiro Ooguchi, Paul Skehel, Tomomi Aida, Junji Takeda, Tomoji Mashimo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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.

Original languageEnglish
Article number318
JournalBMC Genomics
Issue number1
Publication statusPublished - 2 May 2018

Keywords / Materials (for Non-textual outputs)

  • Cre-loxP system
  • CRISPR/Cas9
  • Long single-stranded DNA
  • Zygote electroporation


Dive into the research topics of 'CLICK: One-step generation of conditional knockout mice'. Together they form a unique fingerprint.

Cite this