The CRISPR-Cas13a Gene-Editing System Induces Collateral Cleavage of RNA in Glioma Cells

CRISPR-Cas13a 基因编辑系统诱导胶质瘤细胞中的 RNA 侧面裂解

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作者:Qixue Wang, Xing Liu, Junhu Zhou, Chao Yang, Guangxiu Wang, Yanli Tan, Ye Wu, Sijing Zhang, Kaikai Yi, Chunsheng Kang

Abstract

RNA is rarely used as a therapeutic target due to its flexible structure and instability. CRISPR-Cas13a is a powerful tool for RNA knockdown, and the potential application of CRISPR-Cas13a in cancer cells should be further studied. In this study, overexpression of LwCas13a by lentivirus in glioma cells reveals that crRNA-EGFP induces a "collateral effect" after knocking down the target gene in EGFP-expressing cells. EGFRvIII is a unique EGFR mutant subtype in glioma, and the CRISPR-Cas13a system induces death in EGFRvIII-overexpressing glioma cells. Bulk and single-cell RNA sequencing analysis in U87-Cas13a-EGFRvIII cells confirm the collateral effect of the CRISPR-Cas13a system. Furthermore, CRISPR-Cas13a inhibits the formation of glioma intracranial tumors in mice. The results demonstrate the collateral effect of the CRISPR-Cas13a system in cancer cells and the powerful tumor-eliminating potential of this system.

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