引导编辑系统研究进展

    Recent advances in prime editing system

    • 摘要: 引导编辑(Prime editing,PE)系统是一种全新的、革命性的基因组编辑策略。该系统由引导编辑器(Prime editor)组成,包括nCas9(H840A)与逆转录酶(Reverse transcriptase,RT)的融合蛋白;以及包含PBS(Primer binding site)序列和RT模板(RT template,RTT)序列的pegRNA(Prime editing guide RNA)两大部分。PE系统可以在双链不断裂的情况下实现所有12种类型的碱基替换及小片段DNA增删,是精准编辑的全新范式。自2019年开发至今不到4年时间,PE系统作为一种通用的技术平台,已广泛应用于医疗、农业等各个领域,产生了一大批新种质资源、基因治疗药物等优秀应用案例。PE作为目前最灵活、最具发展前景的基因组精准编辑新手段,仍旧存在效率偏低、大片段操纵能力不足、系统组分设计复杂(如pegRNA)、安全性未全面评估等问题,仍需要深入研究。本文详细介绍了PE系统的技术原理及限制因素,全面总结了PE系统自开发以来的优化策略及在动植物系统、医疗领域的应用现状,并对PE的发展前景进行了展望。

       

      Abstract: Prime editing (PE) system is a newly developed and greatly revolutionized genome editing technology. The system is based on prime editors, which are composed of two components: A fusion protein of nCas9 (H840A) and reverse transcriptase (RT), and a pegRNA which contains a PBS (Primer binding site) sequence and an RT template (RTT) sequence. The PE system can realize all 12 types of base substitutions and small fragment DNA additions and deletions without double-strand breaks, which is a new paradigm for precision editing. In less than 4 years since its development in 2019, the PE system, as a universal technology platform, has been widely used in various fields such as healthcare and agriculture, generating a large number of excellent application cases such as new germplasm resources and gene therapy drugs. PE, as the most flexible and promising new means of precision genome editing, still suffers from low efficiency, insufficient ability to manipulate large fragments, complex design of system components (such as pegRNAs), incomplete evaluation of safety, and still requires in-depth research. This paper described in detail the technical principles and constraints of PE systems, comprehensively summarized the optimization strategies of PE systems since their development, and the current status of PE applications on animal and plant systems and medical fields. It also gave an outlook on the development prospects of PE.

       

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