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

• Main Authors: LIU BIN, DAI LINGLI, BAO YANCHUN, GUO LILI, SHI CAIXIA, ZHANG WENGUANG, LIU ZAIXIA
• Format: Patent
• Language: Chinese; English
• Published: 30.06.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

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