5'-Modifications improve potency and efficacy of DNA donors for precision genome editing.
5'-Modifications improve potency and efficacy of DNA donors for precision genome editing.
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5‘-修饰提高了DNA供体进行精确基因组编辑的效力和效率。
DOI：
10.7554/elife.72216
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发表时间：
2021-10-19
期刊：
影响因子：
7.7
通讯作者：
Mello CC
中科院分区：
文献类型：
--
作者：
Ghanta KS;Chen Z;Mir A;Dokshin GA;Krishnamurthy PM;Yoon Y;Gallant J;Xu P;Zhang XO;Ozturk AR;Shin M;Idrizi F;Liu P;Gneid H;Edraki A;Lawson ND;Rivera-Pérez JA;Sontheimer EJ;Watts JK;Mello CC
核酸酶指导的基因组编辑是研究生理学的强大工具，作为纠正导致疾病的突变的治疗方法有很大的前景。在其最精确的形式中，基因组编辑可以使用细胞同源定向修复(HDR)途径将来自外源提供的DNA修复模板(供体)的信息直接插入到目标基因组位置。不幸的是，尤其是对于长插入，与修复模板DNA相关的毒性和递送考虑可能会限制HDR的疗效。在这里，我们探索双链和单链DNA修复模板的化学修饰。我们描述了5‘末端的修饰，包括以其最简单的形式加入三甘醇(TEG)部分，在三种动物模型(秀丽线虫、斑马鱼、小鼠)的种系和培养的人类细胞中持续提高精确编辑的频率。
Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an exogenously supplied DNA-repair template (donor) directly into a targeted genomic location. Unfortunately, particularly for long insertions, toxicity and delivery considerations associated with repair template DNA can limit HDR efficacy. Here, we explore chemical modifications to both double-stranded and single-stranded DNA-repair templates. We describe 5′-terminal modifications, including in its simplest form the incorporation of triethylene glycol (TEG) moieties, that consistently increase the frequency of precision editing in the germlines of three animal models (Caenorhabditis elegans, zebrafish, mice) and in cultured human cells.