CAS Key Laboratory of Infection and Immunity, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
University of Chinese Academy of Sciences, Beijing, China.
Nat Cell Biol. 2020 Mar;22(3):282-288. doi: 10.1038/s41556-020-0471-6. Epub 2020 Feb 17.
Clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR interference and programmable base editing have transformed the manipulation of eukaryotic genomes for potential therapeutic applications. Here, we exploited CRISPR interference and programmable base editing to determine their potential in editing a TERT gene promoter-activating mutation, which occurs in many diverse cancer types, particularly glioblastoma. Correction of the -124C>T TERT promoter mutation to -124C was achieved using a single guide RNA (sgRNA)-guided and catalytically impaired Campylobacter jejuni CRISPR-associated protein 9-fused adenine base editor (CjABE). This modification blocked the binding of members of the E26 transcription factor family to the TERT promoter, reduced TERT transcription and TERT protein expression, and induced cancer-cell senescence and proliferative arrest. Local injection of adeno-associated viruses expressing sgRNA-guided CjABE inhibited the growth of gliomas harbouring TERT-promoter mutations. These preclinical proof-of-concept studies establish the feasibility of gene editing as a therapeutic approach for cancer and validate activated TERT-promoter mutations as a cancer-specific therapeutic target.
成簇规律间隔短回文重复序列（CRISPR）、CRISPR 干扰和可编程碱基编辑技术已经改变了真核生物基因组的操作，为潜在的治疗应用提供了可能。在这里，我们利用 CRISPR 干扰和可编程碱基编辑来确定它们在编辑 TERT 基因启动子激活突变中的潜力，该突变发生在许多不同的癌症类型中，特别是神经胶质瘤。使用单引导 RNA（sgRNA）引导的和催化失活的空肠弯曲菌 CRISPR 相关蛋白 9 融合腺嘌呤碱基编辑器（CjABE），将-124C>T TERT 启动子突变校正为-124C。这种修饰阻止了 E26 转录因子家族成员与 TERT 启动子的结合，降低了 TERT 转录和 TERT 蛋白表达，并诱导了癌细胞衰老和增殖停滞。表达 sgRNA 引导的 CjABE 的腺相关病毒的局部注射抑制了携带 TERT 启动子突变的神经胶质瘤的生长。这些临床前概念验证研究确立了基因编辑作为癌症治疗方法的可行性，并验证了激活的 TERT 启动子突变作为癌症特异性治疗靶点的可行性。
