中科院化学所汪铭团队报道了正交化学激活酶诱导的CRISPR/Cas9用于细胞选择性基因组编辑。相关研究成果于2022年11月11日发表在《美国化学会杂志》。

CRISPR/Cas9基因组编辑的准确性和治疗潜力因对Cas9介导的DNA切割的较少控制而受到极大挑战。

该文中，研究人员介绍了一种由疾病相关酶控制的条件和细胞选择性基因组编辑系统，称为酶诱导型CRISPR（eiCRISPR）。eiCRISPR包括Cas9蛋白、一种自封闭的非活性单导RNA（bsgRNA）和一种化学笼式脱氧核酶（DNAzyme），其以可控的方式激活bsgRNA和eiCRISPR。研究人员设计了DNAzyme的化学修饰，以抑制其切割bsgRNA阻断区的能力，而由肿瘤细胞过度表达的酶（例如NAD（P）H:醌氧化还原酶（NQO1））触发的DNAzym的去乙酰化，恢复了bsgRNA的活性，并选择性地开启eiCRISPR，用于癌细胞的基因组编辑。

此外，使用可生物降解的脂质纳米粒子在携带肿瘤的异种移植物中递送eiCRISPR，研究表明eiCRISPR的体内激活能够编辑人乳头瘤病毒18 E6以用于潜在的癌症治疗。DNAzyme的合成后和位点特异性修饰策略与内源性化学相容，用于调节eiCRISPR，用于细胞选择性基因组编辑和靶向基因治疗。

附：英文原文

Title: Orthogonal Chemical Activation of Enzyme-Inducible CRISPR/Cas9 for Cell-Selective Genome Editing

Author: Weiqi Cai, Ji Liu, Xianghan Chen, Lanqun Mao, Ming Wang

Issue&Volume: November 11, 2022

Abstract: The precision and therapeutic potential of CRISPR/Cas9 genome editing are greatly challenged by the less control over Cas9-mediated DNA cleavage. Herein, we introduce a conditional and cell-selective genome editing system controlled by disease-associated enzymes, termed enzyme-inducible CRISPR (eiCRISPR). eiCRISPR comprises Cas9 protein, a self-blocked inactive single-guide RNA (bsgRNA), and a chemically caged deoxyribozyme (DNAzyme) that activates bsgRNA and eiCRISPR in a controllable manner. We design chemical modifications of DNAzyme to suppress its ability to cleave the blocking region of bsgRNA, while the decaging of DNAzyme triggered by the tumor cell-overexpressed enzyme, for instance, NAD(P)H:quinone oxidoreductase (NQO1), restores the activity of bsgRNA and switches on eiCRISPR selectively for genome editing in cancer cells. Moreover, using a biodegradable lipid nanoparticle to deliver eiCRISPR in a tumor-bearing xenograft, we show that the in vivo activation of eiCRISPR enables the editing of human papillomavirus 18 E6 for potential cancer therapy. The strategy of postsynthetic and site-specific modification of DNAzyme is compatible with endogenous chemistries for regulating eiCRISPR for cell-selective genome editing and targeted gene therapy.

DOI: 10.1021/jacs.2c10545

Source: https://pubs.acs.org/doi/10.1021/jacs.2c10545