Bovine whole genome-based CRISPR-Cas9 knockout library, knockout cell bank and target gene screening method

The invention provides a bovine whole genome CRISPR-Cas9-based knockout library, a knockout cell bank and a method for screening a target gene, and relates to the technical field of library construction. The invention provides a construction method of a cattle whole genome coding gene CRISPR-Cas9 li...

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Bibliographische Detailangaben
Hauptverfasser: LIU BIN, DAI LINGLI, BAO YANCHUN, GUO LILI, SHI CAIXIA, ZHANG WENGUANG, LIU ZAIXIA
Format: Patent
Sprache:chi ; eng
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Zusammenfassung:The invention provides a bovine whole genome CRISPR-Cas9-based knockout library, a knockout cell bank and a method for screening a target gene, and relates to the technical field of library construction. The invention provides a construction method of a cattle whole genome coding gene CRISPR-Cas9 library, and on the basis of the constructed cattle whole genome CRISPR/Cas9 knockout library, an MDBK cattle kidney epithelial cell whole genome knockout cell library is constructed for the first time. Based on a cattle whole genome CRISPR/Cas9 knockout library strategy, nine candidate genes related to beta-hydroxybutyric acid (BHBA) action are screened and identified for the first time and can be used as molecular markers for drug screening and marker-assisted management or molecular targets for preparing a gene editing disease-resistant animal model. 本发明提供了基于牛全基因组CRISPR-Cas9敲除文库、敲除细胞库及筛选目标基因的方法,涉及文库构建技术领域。本发明提供了一种牛全基因组编码基因CRISPR-Cas9文库的构建方法,并基于构建得到的牛全基因组CRISPR/Cas9敲除文库,首次构建了MDBK牛肾上皮细胞全基因组敲除细胞库。本发明基于牛全基因组CRISPR/Cas