Production of MSTN-mutated cattle without exogenous gene integration using CRISPR-Cas9

被引：29

作者：

Gim, Gyeong-Min ^{[1
,2
,3
]}

Kwon, Dong-Hyeok ^{[1
,2
,3
]}

Eom, Kyeong-Hyun ^{[1
,2
,3
]}

Moon, JoonHo ^{[4
]}

Park, Ji-Hyun ^{[4
]}

Lee, Won-Wu ^{[4
]}

Jung, Dae-Jin ^{[5
]}

Kim, Dae-Hyun ^{[5
]}

Yi, Jun-Koo ^{[5
]}

Ha, Jae-Jung ^{[5
]}

Lim, Ka-Yeong ^{[6
]}

Kim, Jin-Soo ^{[6
]}

Jang, Goo ^{[1
,2
,3
]}

机构：

[1] Seoul Natl Univ, Coll Vet Med, Dept Vet Clin Sci, Lab Theriogenol & Biotechnol, Seoul, South Korea

[2] Seoul Natl Univ, Res Inst Vet Sci, Seoul, South Korea

[3] Seoul Natl Univ, Coll Vet Med, BK21 Plus Program, Seoul, South Korea

[4] LARTBio Inc, Seoul, South Korea

[5] Gyeongsangbukdo Livestock Res Inst, Yeongju, South Korea

[6] Inst Basic Sci IBS, Ctr Genome Engn, Seoul, South Korea

来源：

BIOTECHNOLOGY JOURNAL | 2022年 / 17卷 / 07期

基金：

新加坡国家研究基金会;

关键词：

bovine embryos; CRISPR-Cas9; in vitro fertilization; microinjection; MSTN; MYOSTATIN; MUTATIONS; TOOL;

D O I：

10.1002/biot.202100198

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Many genome-edited animals have been produced using clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology to edit specific genes. However, there are few guidelines for the application of this technique to cattle. The goal of this study was to produce trait-improved cattle using the genome-editing technology CRISPR-Cas9. Myostatin (MSTN) was selected as a target locus, and synthetic mRNA of sgRNA and Cas9 were microinjected into fertilized bovine embryos in vitro. As a result, 17 healthy calves were born, and three of them showed MSTN mutation rates of 10.5%, 45.4%, and 99.9%, respectively. Importantly, the offspring with the 99.9% MSTN mutation rate had a biallelic mutation (-12 bps) and a double-muscling phenotype. In conclusion, we demonstrate that the genome-editing technology CRISPR-Cas9 can produce genetically modified calves with improved traits.

引用

页数：8

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