小柯机器人

韩国首尔延世大学医学院Hyongbum Henry Kim团队近期取得了重要进展，他们利用碱基编辑器对人类癌症相关突变进行高通量功能评估。该项研究成果2022年4月11日在线发表于《自然—生物技术》杂志上。

在这项研究中，研究人员使用胞嘧啶和腺嘌呤碱基编辑器以及单导向RNA（sgRNA）文库，评估了29,060种癌症相关过渡突变（这些突变会导致蛋白质变异）对非致瘤性肺细胞的存活和增殖的功能影响。通过使用与慢病毒递送构建体上的 sgRNA 编码序列配对的替代靶序列监测碱基编辑效率和结果，研究人员确定了sgRNA诱导产生的单个主要蛋白质变体，从而能够将这些突变与碱基编辑引起的细胞表型联系起来。绝大多数蛋白质变体（28,458 个变体，98%）的功能被归类为中性或可能中性；只有18个 (0.06%) 和157个 (0.5%) 变体分别导致生长和可能生长的表型。

研究人员希望他们的方法可以扩展到更多未知意义的变体和其他肿瘤类型中。

据介绍，癌症组织中发现的许多突变的综合表型表征是癌症基因组学中最大的挑战之一。

附：英文原文

Title: High-throughput functional evaluation of human cancer-associated mutations using base editors

Author: Kim, Younggwang, Lee, Seungho, Cho, Soohyuk, Park, Jinman, Chae, Dongwoo, Park, Taeyoung, Minna, John D., Kim, Hyongbum Henry

Issue&Volume: 2022-04-11

Abstract: Comprehensive phenotypic characterization of the many mutations found in cancer tissues is one of the biggest challenges in cancer genomics. In this study, we evaluated the functional effects of 29,060 cancer-related transition mutations that result in protein variants on the survival and proliferation of non-tumorigenic lung cells using cytosine and adenine base editors and single guide RNA (sgRNA) libraries. By monitoring base editing efficiencies and outcomes using surrogate target sequences paired with sgRNA-encoding sequences on the lentiviral delivery construct, we identified sgRNAs that induced a single primary protein variant per sgRNA, enabling linking those mutations to the cellular phenotypes caused by base editing. The functions of the vast majority of the protein variants (28,458 variants, 98%) were classified as neutral or likely neutral; only 18 (0.06%) and 157 (0.5%) variants caused outgrowing and likely outgrowing phenotypes, respectively. We expect that our approach can be extended to more variants of unknown significance and other tumor types. Cancer-associated variants of unknown significance are identified using base editing.

DOI: 10.1038/s41587-022-01276-4

Source: https://www.nature.com/articles/s41587-022-01276-4

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