CRISPR/Cas9-based liver-derived reporter cells for screening of mPGES-1 inhibitors

Journal of Enzyme Inhibition and Medicinal Chemistry

LinLin Yan¹  Luxi Wu¹  Zhanfei Chen¹  Man Li¹  Xiaoqian Wang¹  Xiaoling Cai¹  Nanhong Tang² 

[1] Fujian Institute of Hepatobiliary Surgery, Fujian Medical University Union Hospital, Fuzhou, China;Fujian Institute of Hepatobiliary Surgery, Fujian Medical University Union Hospital, Fuzhou, China;Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Research Center for Molecular Medicine, Fujian Medical University, Fuzhou, Chin;

关键词: CRISPR/Cas9;    liver-derived cells;    mPGES-1;    inhibitor;   

DOI  :  10.1080/14756366.2019.1587416

来源: publisher

【 摘 要 】

mPGES-1 is a terminal rate-limiting enzyme responsible for inflammation-induced PGE2 production. The inhibition of mPGES-1 has been considered as a safe and effective target for the treatment of inflammation and cancer. However, a specific, efficient, and simple method for high-throughput screening of mPGES-1 inhibitors is still lacking. In this study, we developed a fluorescence imaging strategy to monitor the expression of mPGES-1 via CRISPR/Cas9 knock-in system. Immunofluorescence colocalisation, Sanger sequencing, RNAi, and IL-1β treatment all confirmed the successful construction of mPGES-1 reporter cells. The fluorescence signal intensity of the reporter cells treated with four conventional mPGES-1 inhibitors was considerably attenuated via flow cytometry and fluorescent microplate reader, demonstrating that the reporter cells can be used as an efficient and convenient means for screening and optimising mPGES-1 inhibitors. Moreover, it provides a new technical support for the development of targeted small molecule compounds for anti-inflammatory and tumour therapy.

【 授权许可】

CC BY