Zhuang Chengle, Zhuang Changshui, Zhou Qun, Huang Xueting, Gui Yaoting, Lai Yongqing, Yang Shangqi
Department of Urology, Peking University Shenzhen Hospital, Shenzhen, China.
Department of Urology, Union Shenzhen Hospital, Huazhong University of Science and Technology, Shenzhen, China.
Front Mol Biosci. 2021 Mar 19;8:646412. doi: 10.3389/fmolb.2021.646412. eCollection 2021.
Aptazyme and CRISPR/Cas gene editing system were widely used for regulating gene expression in various diseases, including cancer. This work aimed to reconstruct CRISPR/Cas13d tool for sensing hTERT exclusively based on the new device OFF-switch hTERT aptazyme that was inserted into the 3' UTR of the Cas13d. In bladder cancer cells, hTERT ligand bound to aptamer in OFF-switch hTERT aptazyme to inhibit the degradation of Cas13d. Results showed that engineered CRISPR/Cas13d sensing hTERT suppressed cell proliferation, migration, invasion and induced cell apoptosis in bladder cancer 5637 and T24 cells without affecting normal HFF cells. In short, we constructed engineered CRISPR/Cas13d sensing hTERT selectively inhibited the progression of bladder cancer cells significantly. It may serve as a promising specifically effective therapy for bladder cancer cells.
适体酶和CRISPR/Cas基因编辑系统被广泛用于调控包括癌症在内的各种疾病中的基因表达。这项工作旨在基于插入到Cas13d的3' UTR中的新型关闭开关hTERT适体酶,重建专门用于检测hTERT的CRISPR/Cas13d工具。在膀胱癌细胞中,hTERT配体与关闭开关hTERT适体酶中的适体结合,以抑制Cas13d的降解。结果表明,工程化的CRISPR/Cas13d检测hTERT可抑制膀胱癌5637和T24细胞的增殖、迁移、侵袭并诱导细胞凋亡,而不影响正常的HFF细胞。简而言之,我们构建的工程化CRISPR/Cas13d检测hTERT可选择性地显著抑制膀胱癌细胞的进展。它可能成为一种有前景的针对膀胱癌细胞的特异性有效疗法。