基础医学与临床2025，Vol.45Issue(8)：1016-1021,6.DOI:10.16352/j.issn.1001-6325.2025.08.1016
用CRISPR/Cas 9技术构建ACSS3基因稳定敲除的肺癌细胞株
Stable knockout of ACSS3 in lung cancer cell line using CRISPR/Cas 9 technology
摘要
Abstract
Objective To explore the effect of acyl-CoA synthetase short-chain family member 3(ACSS3)gene on the proliferation of human large cell lung cancer cells(NCI-H460)using CRISPR/Cas 9 gene editing technology.Methods The expression of ACSS3 was detected by Western blot.ACSS3-targeting sgRNAs were designed,and a CRISPR/Cas 9 knockout vector was constructed and transfected into NCI-H460 cells.The transfected cells were selected with puromycin based on vector-carried resistance.ACSS3-knockout monoclonal cell strains were established by limited dilution method and then expanded in culture.Knockout efficiency was confirmed by Western blot.Cell proliferation was assessed using MTT and colony formation assays.Results The expression of ACSS3 was significantly elevated in NCI-H460 cells as compared with human normal lung epithelial cells BEAS-2B(P＜0.05).No ACSS3 protein was detected in the knockout monoclonal strain,indicating successful generation of ACSS3-knockout NCI-H460 cells.Compared with the control cells transfected with empty vector,the proliferation and colo-ny formation ability were inhibited in NCI-H460 cells with ACSS3 knockout(P＜0.05).Conclusions The ACSS3-knockout NCI-H460 cell strain was successfully established,which provides a foundation for further study on the role of ACSS3 in lung cancer.关键词
肺癌/CRISPR/Cas 9/ACSS3Key words
lung cancer/CRISPR/Cas 9/ACSS3分类
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黄倩倩,贾玉芳,余华军,陈融融,陈丽丽,伍俊,张海涛..用CRISPR/Cas 9技术构建ACSS3基因稳定敲除的肺癌细胞株[J].基础医学与临床,2025,45(8):1016-1021,6.基金项目
广东省基础与应用基础研究基金自然科学基金项目(2024A1515013157) （2024A1515013157）
湛江市科技发展专项资金竞争性分配项目(2022A702-1) （2022A702-1）
