Stable knockout of ACSS3 in lung cancer cell line using CRISPR/Cas 9 technology
10.16352/j.issn.1001-6325.2025.08.1016
- VernacularTitle:用CRISPR/Cas 9技术构建ACSS3基因稳定敲除的肺癌细胞株
- Author:
Qianqian HUANG
1
;
Yufang JIA
;
Huajun YU
;
Rongrong CHEN
;
Lili CHEN
;
Jun WU
;
Haitao ZHANG
Author Information
1. 广东医科大学医学技术学院,广东湛江 524023;广东医科大学生物化学与分子生物学研究所&多肽和蛋白研究与应用重点实验室,广东湛江 524023
- Keywords:
lung cancer;
CRISPR/Cas 9;
ACSS3
- From:
Basic & Clinical Medicine
2025;45(8):1016-1021
- CountryChina
- Language:Chinese
-
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.
