Wang Qixue, Liu Xing, Zhou Junhu, Yang Chao, Wang Guangxiu, Tan Yanli, Wu Ye, Zhang Sijing, Yi Kaikai, Kang Chunsheng
Tianjin Medical University General Hospital Tianjin Neurological Institute Key Laboratory of Post-neurotrauma Neuro-repair and Regeneration in Central Nervous System Ministry of Education and Tianjin City Department of Neurosurgery Tianjin Medical University General Hospital Tianjin 300052 China.
Beijing Neurosurgical Institute Capital Medical University Beijing 100050 China.
Adv Sci (Weinh). 2019 Aug 29;6(20):1901299. doi: 10.1002/advs.201901299. eCollection 2019 Oct 16.
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.
由于RNA结构灵活且不稳定，它很少被用作治疗靶点。CRISPR-Cas13a是一种强大的RNA敲低工具，其在癌细胞中的潜在应用值得进一步研究。在本研究中，通过慢病毒在胶质瘤细胞中过表达LwCas13a发现，crRNA-EGFP在敲低表达EGFP细胞中的靶基因后会诱导“附带效应”。EGFRvIII是胶质瘤中一种独特的EGFR突变亚型，CRISPR-Cas13a系统可诱导EGFRvIII过表达的胶质瘤细胞死亡。对U87-Cas13a-EGFRvIII细胞进行的批量和单细胞RNA测序分析证实了CRISPR-Cas13a系统的附带效应。此外，CRISPR-Cas13a可抑制小鼠颅内胶质瘤肿瘤的形成。结果证明了CRISPR-Cas13a系统在癌细胞中的附带效应以及该系统强大的肿瘤消除潜力。
