[HTML][HTML] Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice
T Aida, K Chiyo, T Usami, H Ishikubo, R Imahashi… - Genome biology, 2015 - Springer
Genome biology, 2015•Springer
Although the CRISPR/Cas system has enabled one-step generation of knockout mice, low
success rates of cassette knock-in limit its application range. Here we show that cloning-free,
direct nuclear delivery of Cas9 protein complex with chemically synthesized dual RNAs
enables highly efficient target digestion, leading to generation of knock-in mice carrying a
functional cassette with up to 50% efficiency, compared with just 10% by a commonly used
method consisting of Cas9 mRNA and single guide RNA. Our cloning-free CRISPR/Cas …
success rates of cassette knock-in limit its application range. Here we show that cloning-free,
direct nuclear delivery of Cas9 protein complex with chemically synthesized dual RNAs
enables highly efficient target digestion, leading to generation of knock-in mice carrying a
functional cassette with up to 50% efficiency, compared with just 10% by a commonly used
method consisting of Cas9 mRNA and single guide RNA. Our cloning-free CRISPR/Cas …
Abstract
Although the CRISPR/Cas system has enabled one-step generation of knockout mice, low success rates of cassette knock-in limit its application range. Here we show that cloning-free, direct nuclear delivery of Cas9 protein complex with chemically synthesized dual RNAs enables highly efficient target digestion, leading to generation of knock-in mice carrying a functional cassette with up to 50% efficiency, compared with just 10% by a commonly used method consisting of Cas9 mRNA and single guide RNA. Our cloning-free CRISPR/Cas system facilitates rapid one-step generation of cassette knock-in mice, accelerating functional genomic research by providing various in vivo genetic tools.
Springer