小柯机器人

美国华盛顿大学Jay Shendure、Junhong Choi等研究人员合作开发出能够进行精确基因组删除的新工具。这一研究成果于2021年10月14日在线发表在国际学术期刊《自然—生物技术》上。

研究人员描述了一种基于prime editing的方法——PRIME-Del，它使用一对prime editing sgRNA（pegRNA）来诱导缺失，这些sgRNA的目标是相反的DNA链，不仅对被缺口的位点进行编程，也对修复的结果进行编程。与CRISPR-Cas9和sgRNA对相比，PRIME-Del在对长达10kb的缺失进行编程时实现了明显的高精确度，编辑效率为1-30%。PRIME-Del还可用于将基因组缺失与短的插入结合起来，使缺失的结点不落在原型间隔序列-相邻模体位点。最后，在不影响精确性的前提下，扩展表达prime editing元件可以大幅提高效率。研究人员认为PRIME-Del将广泛用于精确、灵活的基因组缺失编程、表位标记以及潜在的基因组重排编程。

据悉，目前删除基因组序列的方法是基于CRISPR-Cas9和sgRNA，但这可能是低效和不精确的，出现的错误包括小的插入缺失以及无意的大片段缺失和更复杂的重排。

附：英文原文

Title: Precise genomic deletions using paired prime editing

Author: Choi, Junhong, Chen, Wei, Suiter, Chase C., Lee, Choli, Chardon, Florence M., Yang, Wei, Leith, Anh, Daza, Riza M., Martin, Beth, Shendure, Jay

Issue&Volume: 2021-10-14

Abstract: Current methods to delete genomic sequences are based on clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 and pairs of single-guide RNAs (sgRNAs), but can be inefficient and imprecise, with errors including small indels as well as unintended large deletions and more complex rearrangements. In the present study, we describe a prime editing-based method, PRIME-Del, which induces a deletion using a pair of prime editing sgRNAs (pegRNAs) that target opposite DNA strands, programming not only the sites that are nicked but also the outcome of the repair. PRIME-Del achieves markedly higher precision than CRISPR–Cas9 and sgRNA pairs in programming deletions up to 10kb, with 1–30% editing efficiency. PRIME-Del can also be used to couple genomic deletions with short insertions, enabling deletions with junctions that do not fall at protospacer-adjacent motif sites. Finally, extended expression of prime editing components can substantially enhance efficiency without compromising precision. We anticipate that PRIME-Del will be broadly useful for precise, flexible programming of genomic deletions, epitope tagging and, potentially, programming genomic rearrangements.

DOI: 10.1038/s41587-021-01025-z

Source: https://www.nature.com/articles/s41587-021-01025-z

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex