Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP
文献类型：期刊论文
| 作者 | Bi, Yanzhen1; Hua, Zaidong1; Liu, Ximei1; Hua, Wenjun1; Ren, Hongyan1; Xiao, Hongwei1; Zhang, Liping1; Li, Li1; Wang, Zhirui2; Laible, Gotz3 |
| 刊名 | SCIENTIFIC REPORTS
 |
| 出版日期 | 2016-08-17 |
| 卷号 | 6 |
| ISSN号 | 2045-2322 |
| 英文摘要 | Predictable, clean genetic modification (GM) in livestock is important for reliable phenotyping and biosafety. Here we reported the generation of isozygous, functional myostatin (MSTN) knockout cloned pigs free of selectable marker gene (SMG) by CRISPR/Cas9 and Cre/LoxP. CRISPR/Cas9-mediated homologous recombination (HR) was exploited to knock out (KO) one allele of MSTN in pig primary cells. Cre recombinase was then used to excise the SMG with an efficiency of 82.7%. The SMG-free non-EGFP cells were isolated by flow cytometery and immediately used as donor nuclei for nuclear transfer. A total of 685 reconstructed embryos were transferred into three surrogates with one delivering two male live piglets. Molecular testing verified the mono-allelic MSTN KO and SMG deletion in these cloned pigs. Western blots showed approximately 50% decrease in MSTN and concurrent increased expression of myogenic genes in muscle. Histological examination revealed the enhanced myofiber quantity but myofiber size remained unaltered. Ultrasonic detection showed the increased longissimus muscle size and decreased backfat thickness. Precision editing of pig MSTN gene has generated isozygous, SMG-free MSTN KO cloned founders, which guaranteed a reliable route for elite livestock production and a strategy to minimize potential biological risks. |
| WOS标题词 | Science & Technology |
| 类目[WOS] | Multidisciplinary Sciences |
| 研究领域[WOS] | Science & Technology - Other Topics |
| 关键词[WOS] | ZINC-FINGER NUCLEASES ; DOUBLE-MUSCLED PHENOTYPE ; EMBRYONIC STEM-CELLS ; MYOSTATIN GENE ; CRE RECOMBINASE ; EFFICIENT GENERATION ; SKELETAL-MUSCLE ; BOVINE GENOME ; CATTLE ; EXPRESSION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000381480800001 |
| 源URL | [http://ir.ihb.ac.cn/handle/342005/28334]  |
| 专题 | 水生生物研究所_藻类生物学及应用研究中心_学位论文 |
| 作者单位 | 1.Hubei Acad AgroSci, Hubei Inst Anim Sci & Vet Med, Hubei Key Lab Anim Embryo Engn & Mol Breeding, Wuhan 430064, Peoples R China 2.Henan Univ Sci & Technol, Coll Anim Sci, 263 Kaiyuan Ave, Luoyang 471023, Peoples R China 3.AgResearch, Ruakura Res Ctr, Private Bag 3123, Hamilton, New Zealand 4.Chinese Acad Sci, Inst Hydrobiol, Anal & Testing Ctr, Wuhan 430072, Peoples R China |
| 推荐引用方式 GB/T 7714 | Bi, Yanzhen,Hua, Zaidong,Liu, Ximei,et al. Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP[J]. SCIENTIFIC REPORTS,2016,6. |
| APA | Bi, Yanzhen.,Hua, Zaidong.,Liu, Ximei.,Hua, Wenjun.,Ren, Hongyan.,...&Zheng, Xinmin.(2016).Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP.SCIENTIFIC REPORTS,6. |
| MLA | Bi, Yanzhen,et al."Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP".SCIENTIFIC REPORTS 6(2016). |
入库方式： OAI收割
来源：水生生物研究所
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