Journal
BMC GENOMICS
Volume 19, Issue -, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/s12864-018-4713-y
Keywords
CRISPR/Cas9; CLICK; Zygote electroporation; Long single-stranded DNA; Cre-loxP system
Funding
- Japan Society for the Promotion of Science [26290033, 16 K07085, 26830131]
- CNSI/NINS
- MRI/TMDU
- Nakatani Foundation
- SEN-SHIN Medical Research Foundation
- Mochida Memorial Foundation
- Takeda Science Foundation
- Grants-in-Aid for Scientific Research [26830131, 26290033] Funding Source: KAKEN
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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.
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