CRISPR/Cas9

被引：4

作者：

Mizuno, Naoaki ^{[1
]}

Mizutani, Eiji ^{[1
]}

Sato, Hideyuki ^{[1
]}

Kasai, Mariko ^{[1
]}

Nakauchi, Hiromitsu ^{[1
,2
]}

Yamaguchi, Tomoyuki ^{[1
]}

机构：

[1] Univ Tokyo, Inst Med Sci, Div Stem Cell Therapy, Minato Ku, Tokyo, Japan

[2] Stanford Univ, Sch Med, Dept Genet, Inst Stem Cell Biol & Regenerat Med, Stanford, CA USA

来源：

BIO-PROTOCOL | 2019年 / 9卷 / 13期

关键词：

CRISPR/Cas9; Adeno-associated viral vector; Trans-zona pellucida; Intra-embryo genome editing; Ribonucleoprotein electroporation; Large fragment knock-in; ELECTROPORATION;

D O I：

10.21769/BioProtoc.3295

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Intra-embryo genome editing by CRISPR/Cas9 has enabled rapid generation of gene knockout animals. However, large fragment knock-in directly into embryos' genome is still difficult, especially without microinjection of donor DNA. Viral vectors are good transporters of knock-in donor DNA for cell lines, but seemed unsuitable for pre-implantation embryos with zona pellucida, glycoprotein membrane surrounding early embryos. We found adeno-associated virus (AAV) can infect zygotes of various mammals through intact zona pellucida. AAV-mediated donor DNA delivery following Cas9 ribonucleoprotein electroporation enables large fragment knock-in without micromanipulation.

引用

页数：10

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