Highly efficient RNA-guided base editing in mouse embryos

被引：331

作者：

Kim, Kyoungmi ^{[1
]}

Ryu, Seuk-Min ^{[1
,2
]}

Kim, Sang-Tae ^{[1
]}

Baek, Gayoung ^{[1
]}

Kim, Daesik ^{[2
]}

Lim, Kayeong ^{[1
,2
]}

Chung, Eugene ^{[1
,2
]}

Kim, Sunghyun ^{[1
,2
]}

Kim, Jin-Soo ^{[1
,2
]}

机构：

[1] Inst for Basic Sci Korea, Ctr Genome Engn, Seoul, South Korea

[2] Seoul Natl Univ, Dept Chem, Seoul, South Korea

来源：

NATURE BIOTECHNOLOGY | 2017年 / 35卷 / 05期

关键词：

HUMAN-CELLS; CAS9; ENDONUCLEASES; MICE; SPECIFICITY; MUTAGENESIS; NICKASES; KNOCKOUT;

D O I：

10.1038/nbt.3816

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Base editors (BEs) composed of a cytidine deaminase fused to CRISPR-Cas9 convert cytidine to uridine, leading to single-base-pair substitutions in eukaryotic cells. We delivered BE mRNA or ribonucleoproteins targeting the Dmd or Tyr gene via electroporation or microinjection into mouse zygotes. F0 mice showed nonsense mutations with an efficiency of 44-57% and allelic frequencies of up to 100%, demonstrating an efficient method to generate mice with targeted point mutations.

引用

页码：435 / +

页数：4

共 15 条

[1] Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases [J].

Bae, Sangsu ;

Park, Jeongbin ;

Kim, Jin-Soo .

BIOINFORMATICS, 2014, 30 (10) :1473-1475

[2] Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases [J].

Cho, Seung Woo ;

Kim, Sojung ;

Kim, Yongsub ;

Kweon, Jiyeon ;

Kim, Heon Seok ;

Bae, Sangsu ;

Kim, Jin-Soo .

GENOME RESEARCH, 2014, 24 (01) :132-141

[3] Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease [J].

Cho, Seung Woo ;

Kim, Sojung ;

Kim, Jong Min ;

Kim, Jin-Soo .

NATURE BIOTECHNOLOGY, 2013, 31 (03) :230-232

[4] The AID/APOBEC family of nucleic acid mutators [J].

Conticello, Silvestro G. .

GENOME BIOLOGY, 2008, 9 (06)

[5] Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins [J].

Hur, Junho K. ;

Kim, Kyoungmi ;

Been, Kyung Wook ;

Baek, Gayoung ;

Ye, Sunghyeok ;

Hur, Junseok W. ;

Ryu, Seuk-Min ;

Lee, Youn Su ;

Kim, Jin-Soo .

NATURE BIOTECHNOLOGY, 2016, 34 (08) :807-808

[6]

Kim D, 2015, NAT METHODS, V12, P237, DOI [10.1038/NMETH.3284, 10.1038/nmeth.3284]

[7] Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage [J].

Komor, Alexis C. ;

Kim, Yongjoo B. ;

Packer, Michael S. ;

Zuris, John A. ;

Liu, David R. .

NATURE, 2016, 533 (7603) :420-+

[8]

Li J, 2016, MOL PLANT

[9]

Lu Y., MOL PLANT

[10]

Ma YQ, 2016, NAT METHODS, V13, P1029, DOI [10.1038/NMETH.4027, 10.1038/nmeth.4027]

← 1 2 →