Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated enzyme (Cas) is a naturally occurring genome editing tool adopted from the prokaryotic adaptive immune defense system. Currently, CRISPR/Cas9‐based genome editing has been becoming one of the most promising tools for treating human genetic diseases, including cardiovascular diseases, neuro‐disorders, and cancers. As the quick modification of the CRISPR/Cas9 system, including delivery system, CRISPR/Cas9‐based gene therapy has been extensively studied in preclinic and clinic treatments. CRISPR/Cas genome editing is also a robust tool to create animal genetic models for studying and treating human genetic disorders, particularly diseases associated with point mutations. However, significant challenges also remain before CRISPR/Cas technology can be routinely employed in the clinic for treating different genetic diseases, which include toxicity and immune response of treated cells to CRISPR/Cas component, highly throughput delivery method, and potential off‐target impact. The off‐target effect is one of the major concerns for CRISPR/Cas9 gene therapy, more research should be focused on limiting this impact by designing high specific gRNAs and using high specificity of Cas enzymes. Modifying the CRISPR/Cas9 delivery method not only targets a specific tissue/cell but also potentially limits the off‐target impact.

中文翻译：

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CRISPR/Cas基因治疗

成簇的规则间隔短回文重复序列 (CRISPR) / CRISPR 相关酶 (Cas) 是一种从原核适应性免疫防御系统中采用的天然基因组编辑工具。目前，基于 CRISPR/Cas9 的基因组编辑已成为治疗人类遗传疾病（包括心血管疾病、神经疾病和癌症）最有前途的工具之一。作为包括递送系统在内的 CRISPR/Cas9 系统的快速修改，基于 CRISPR/Cas9 的基因治疗已在临床前和临床治疗中得到广泛研究。CRISPR/Cas 基因组编辑也是创建用于研究和治疗人类遗传疾病，尤其是与点突变相关疾病的动物遗传模型的强大工具。然而，在 CRISPR/Cas 技术在临床上常规用于治疗不同的遗传疾病之前，仍然存在重大挑战，包括治疗细胞对 CRISPR/Cas 成分的毒性和免疫反应、高通量递送方法和潜在的脱靶影响。脱靶效应是 CRISPR/Cas9 基因治疗的主要关注点之一，更多的研究应该集中在通过设计高特异性 gRNA 和使用高特异性 Cas 酶来限制这种影响。修改 CRISPR/Cas9 递送方法不仅针对特定组织/细胞，而且还可能限制脱靶影响。脱靶效应是 CRISPR/Cas9 基因治疗的主要问题之一，更多的研究应该集中在通过设计高特异性 gRNA 和使用高特异性 Cas 酶来限制这种影响。修改 CRISPR/Cas9 递送方法不仅针对特定组织/细胞，而且还可能限制脱靶影响。脱靶效应是 CRISPR/Cas9 基因治疗的主要问题之一，更多的研究应该集中在通过设计高特异性 gRNA 和使用高特异性 Cas 酶来限制这种影响。修改 CRISPR/Cas9 递送方法不仅针对特定组织/细胞，而且还可能限制脱靶影响。