Zygote-mediated generation of genome-modified mice using Streptococcus thermophilus 1-derived CRISPR/Cas system.
Mammalian zygote-mediated genome-engineering by CRISPR/Cas is currently used for the generation of genome-modified animals. Here we report that a Streptococcus thermophilus-1 derived orthologous CRISPR/Cas system, which recognizes the 5'-NNAGAA sequence as a protospacer adjacent motif (PAM), is useful in mouse zygotes and is applicable for generating knockout mice (87.5%) and targeted knock-in mice (45.5%). The induced mutation could be inherited in the next generation. This novel CRISPR/Cas can expand the feasibility of the zygote-mediated generation of genome-modified animals that require an exact mutation design.
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan. Electronic address: awtrfj@mail.ecc.u-tokyo.ac.jp.
Wataru
W
Fujii
Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
Shigeru
S
Kakuta
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
Shin
S
Yoshioka
Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
Shigeru
S
Kyuwa
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
Koji
K
Sugiura
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan. Electronic address: aknaito@mail.ecc.u-tokyo.ac.jp.
Kunihiko
K
Naito
D000818:Animals; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D005260:Female; D016678:Genome; D051379:Mice; D008810:Mice, Inbred C57BL; D013291:Streptococcus / Q000378:metabolism*; D015053:Zygote / Q000378:metabolism*
CRISPR/Cas; Genome engineering; Knockin mice; Knockout mice