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基于 CRISPR 的基因组编辑工具的研究进展综述:碱基编辑和先导编辑。

A Review on Advanced CRISPR-Based Genome-Editing Tools: Base Editing and Prime Editing.

机构信息

Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Mol Biotechnol. 2023 Jun;65(6):849-860. doi: 10.1007/s12033-022-00639-1. Epub 2022 Dec 22.

Abstract

In the field of medicine, it is axiomatic that the need of a precise gene-editing tool is critical to employ therapeutic approaches toward pathogenic mutations, occurring in human genome. Today we know that most of genetic defects are caused by single-base pair substitutions in genomic DNA. The ability to make practically any targeted substitutions of DNA sequences at specified regions in the human genome gives us the chance to employ gene therapy in most known diseases associated with genetic variants. In this regard, CRISPR/Cas9 applications is becoming more and more popular along with the significant advancements of life sciences, by employing this technology in genome-editing and high-throughput screenings. Several CRISPR/Cas-based mammalian cell gene-editing techniques have been developed during the last decade, including nucleases, base editors, and prime editors, all of which have the exact mechanism at first glance. However, they address a subset of known pathogenic sequence mutations using different methods. First, we highlight the development of CRISPR-based gene-editing tools. Then we describe their functions and summarize the conducted research studies, which are increasing the reliability of these strategies to better efficiencies for prospective gene therapies in the near future. Lastly, we compare the capabilities of all these platforms together besides their probable limitations.

摘要

在医学领域,有一个公理,即需要一种精确的基因编辑工具,这对于采用治疗方法来处理人类基因组中发生的致病突变至关重要。如今,我们知道大多数遗传缺陷是由基因组 DNA 中的单个碱基对替换引起的。能够在人类基因组的特定区域对 DNA 序列进行几乎任何靶向替换,这为我们提供了在大多数与遗传变异相关的已知疾病中采用基因治疗的机会。在这方面,随着生命科学的显著进步,CRISPR/Cas9 的应用越来越受欢迎,通过在基因组编辑和高通量筛选中使用这项技术。在过去十年中,已经开发了几种基于 CRISPR 的哺乳动物细胞基因编辑技术,包括核酸酶、碱基编辑器和 Prime 编辑器,它们的精确机制乍一看是相同的。然而,它们使用不同的方法来解决已知致病序列突变的一个子集。首先,我们强调了基于 CRISPR 的基因编辑工具的发展。然后我们描述了它们的功能,并总结了正在进行的研究,这些研究提高了这些策略的可靠性,以便在不久的将来更好地提高潜在基因治疗的效率。最后,我们比较了所有这些平台的能力,以及它们可能存在的局限性。

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