美国马萨诸塞州总医院Benjamin P. Kleinstiver课题组发现，利用DNA聚合酶和HUH内切酶进行点击编辑实现可编程基因组编写。相关论文于2024年7月22日在线发表于国际学术期刊《自然—生物技术》。

研究人员开发了一种名为点击编辑的基因书写平台，该平台将DNA依赖性聚合酶（DDP）的优良特性与RNA可编程切口酶结合，允许进行替换、插入和删除等多种编辑。点击编辑器（CE）利用HUH核酸内切酶与单链DNA底物的“点击”式生物偶联能力，将编码用户指定编辑的“点击DNA”（clkDNA）模板共价连接到目标基因组位点。

通过对CE及其clkDNA模块组件的迭代优化，研究人员展示了在多种永生化人细胞类型和原代成纤维细胞中以高达约30%的精确编辑效率进行精准基因组编辑且产生的插入缺失（indel）极少。通过快速筛选具有各种修饰（包括修复规避替换）的clkDNA寡核苷酸，可以进一步提高编辑效率。点击编辑是一种精确且多功能的基因编辑方法，适用于多种生物学应用。

研究人员表示，基于DDP的基因编辑技术相比其他类型的编辑器具有多种优势，可用于进行多种编辑。

附：英文原文

Title: Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases

Author: Ferreira da Silva, Joana, Tou, Connor J., King, Emily M., Eller, Madeline L., Rufino-Ramos, David, Ma, Linyuan, Cromwell, Christopher R., Metovic, Jasna, Benning, Friederike M. C., Chao, Luke H., Eichler, Florian S., Kleinstiver, Benjamin P.

Issue&Volume: 2024-07-22

Abstract: Genome editing technologies based on DNA-dependent polymerases (DDPs) could offer several benefits compared with other types of editors to install diverse edits. Here, we develop click editing, a genome writing platform that couples the advantageous properties of DDPs with RNA-programmable nickases to permit the installation of a range of edits, including substitutions, insertions and deletions. Click editors (CEs) leverage the ‘click’-like bioconjugation ability of HUH endonucleases with single-stranded DNA substrates to covalently tether ‘click DNA’ (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs and their clkDNAs, we demonstrate the ability to install precise genome edits with minimal indels in diverse immortalized human cell types and primary fibroblasts with precise editing efficiencies of up to ~30%. Editing efficiency can be improved by rapidly screening clkDNA oligonucleotides with various modifications, including repair-evading substitutions. Click editing is a precise and versatile genome editing approach for diverse biological applications.

DOI: 10.1038/s41587-024-02324-x

Source: https://www.nature.com/articles/s41587-024-02324-x