Laboratory of Genome Editing, Research Center for Medical Genetics, Moskvorechye 1, 115478 Moscow, Russia.
Int J Mol Sci. 2023 Dec 1;24(23):17045. doi: 10.3390/ijms242317045.
Prime editing is a rapidly developing method of CRISPR/Cas-based genome editing. The increasing number of novel PE applications and improved versions demands constant analysis and evaluation. The present review covers the mechanism of prime editing, the optimization of the method and the possible next step in the evolution of CRISPR/Cas9-associated genome editing. The basic components of a prime editing system are a prime editor fusion protein, consisting of nickase and reverse transcriptase, and prime editing guide RNA, consisting of a protospacer, scaffold, primer binding site and reverse transcription template. Some prime editing systems include other parts, such as additional RNA molecules. All of these components were optimized to achieve better efficiency for different target organisms and/or compactization for viral delivery. Insights into prime editing mechanisms allowed us to increase the efficiency by recruiting mismatch repair inhibitors. However, the next step in prime editing evolution requires the incorporation of new mechanisms. Prime editors combined with integrases allow us to combine the precision of prime editing with the target insertion of large, several-kilobase-long DNA fragments.
碱基编辑是一种基于 CRISPR/Cas 的快速发展的基因组编辑方法。越来越多的新型 PE 应用和改进版本要求不断进行分析和评估。本综述涵盖了碱基编辑的机制、方法的优化以及 CRISPR/Cas9 相关基因组编辑的可能的下一步发展。碱基编辑系统的基本组成部分是由核酸酶和逆转录酶组成的碱基编辑融合蛋白，以及由原间隔序列、支架、引物结合位点和逆转录模板组成的碱基编辑指导 RNA。一些碱基编辑系统包括其他部分，如额外的 RNA 分子。所有这些成分都经过优化，以提高不同靶标生物的效率和/或简化病毒传递。对碱基编辑机制的深入了解使我们能够通过招募错配修复抑制剂来提高效率。然而，碱基编辑进化的下一步需要引入新的机制。将整合酶与碱基编辑器结合，使我们能够将碱基编辑的精确性与几 kb 长的大型 DNA 片段的靶向插入相结合。
