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CLICK: one-step generation of conditional knockout mice
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DOI：10.1186/s12864-018-4713-y.png)
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Abstract
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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.
Keywords:
CRISPR/Cas9
CLICK
Zygote electroporation
Long single-stranded DNA
Cre-loxP system
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