Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing.
The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing.
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Linlin
L
Zhang
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Jiankui
J
Zhou
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Jinxiong
J
Han
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Bian
B
Hu
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Ningning
N
Hou
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Yun
Y
Shi
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.|School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
Xingxu
X
Huang
Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China.
Xin
0000-0002-6715-0375
X
Lou
D000483:Alleles; D000818:Animals; D064113:CRISPR-Cas Systems; D017931:DNA Primers / Q000235:genetics; D005260:Female; D000072669:Gene Editing; D018390:Gene Targeting; D016678:Genome; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016296:Mutagenesis; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000235:genetics; D009865:Oocytes / Q000166:cytology*; D016133:Polymerase Chain Reaction; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics; D017422:Sequence Analysis, DNA; D000072216:Zona Pellucida Glycoproteins / Q000235:genetics
C000607144:AR protein, mouse; D017931:DNA Primers; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D000094704:RNA, Guide, CRISPR-Cas Systems; D011944:Receptors, Androgen; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse