鉴定猪 Y 染色体上基因的安全港位点。

Identification of Site in the Gene as Safe Harbor Locus on the Y Chromosome of Pig.

机构信息

State Key Laboratory of Farm Animal Biotech Breeding, Frontiers Science Center for Molecular Design Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.

College of Animal Science and Technology, Sanya Institute of China Agricultural University, Sanya 572025, China.

出版信息

Genes (Basel). 2024 Aug 1;15(8):1005. doi: 10.3390/genes15081005.

DOI:10.3390/genes15081005

PMID:39202365

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11353466/

Abstract

Genomic Safe Harbors (GSH) are loci used for the insertion of exogenous genetic elements, enabling exogenous gene expressing predictably without alterations of the host genome. These sites are becoming increasingly important as the gene editing technologies advance rapidly. Currently, only a few GSHs have been identified in the pig genome. In this study, a novel strategy was demonstrated for the efficient insertion of exogenous genetic material into the third exon of the gene on the Y chromosome using CRISPR/Cas9-mediated homology arm-mediated end joining. The safety of the locus was verified according to the proper expression of the inserted gene without altering the expression of . This approach enables the integration and expression of the exogenous gene at this locus, indicating that the locus serves as a genomic safe harbor site for gene editing in the pig genome. Located on the Y chromosome, this site can be utilized for sex-biased pig breeding and developing biomedical models.

摘要

基因组安全港（Genomic Safe Harbors，GSH）是用于插入外源遗传元件的位点，能够在外源基因表达可预测而不改变宿主基因组的情况下实现。随着基因编辑技术的快速发展，这些位点变得越来越重要。目前，在猪基因组中仅鉴定出少数几个 GSH。在这项研究中，展示了一种新的策略，用于使用 CRISPR/Cas9 介导的同源臂介导的末端连接，在 Y 染色体上的基因的第三个外显子中高效插入外源遗传物质。根据插入的基因的正确表达而没有改变的表达，验证了该位点的安全性。这种方法能够使外源基因在该位点进行整合和表达，表明位点是猪基因组基因编辑的基因组安全港位点。该位点位于 Y 染色体上，可用于性别偏向性猪繁殖和开发生物医学模型。