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鉴定基因作为猪基因组中潜在的肌肉特异性安全港基因座。

Identification of the Gene as a Potential Muscle-Specific Safe Harbor Locus in Pig Genome.

机构信息

Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, Key Laboratory of Animal Molecular Design and Precise Breeding of Guangdong Higher Education Institutes, School of Life Science and Engineering, Foshan University, Foshan 528225, China.

Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, China.

出版信息

Genes (Basel). 2022 May 21;13(5):921. doi: 10.3390/genes13050921.

Abstract

Genetically modified pigs have shown considerable application potential in the fields of life science research and livestock breeding. Nevertheless, a barrier impedes the production of genetically modified pigs. There are too few safe harbor loci for the insertion of foreign genes into the pig genome. Only a few loci (, and ) have been successfully identified to achieve the ectopic expression of foreign genes and produce gene-edited pigs. Here, we use CRISPR/Cas9-mediated homologous directed repair (HDR) to accurately knock the exogenous gene-of-interest fragments into an endogenous gene in the porcine satellite cells. After porcine satellite cells are induced to differentiate, the gene promoter simultaneously initiates the expression of the gene and the exogenous gene. We infer preliminarily that the gene can be identified as a potential muscle-specific safe harbor locus in pigs for the integration of exogenous gene-of-interest fragments.

摘要

基因修饰猪在生命科学研究和家畜育种领域显示出了相当大的应用潜力。然而,有一个障碍阻碍了基因修饰猪的生产。将外源基因插入猪基因组的安全港基因座太少。只有少数几个基因座(,和)已被成功鉴定为实现外源基因的异位表达,并产生基因编辑猪。在这里,我们使用 CRISPR/Cas9 介导的同源定向修复(HDR)技术,将目的基因片段精确地敲入猪卫星细胞中的内源性基因中。在猪卫星细胞被诱导分化后,该基因启动子同时启动目的基因和外源基因的表达。我们初步推断,该基因可以作为猪中外源基因片段整合的潜在肌肉特异性安全港基因座。

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