小柯机器人

丹麦哥本哈根大学Morten Frödin和Claus Storgaard Sørensen研究团队合作的最新研究利用CRISPR-Select对基因序列变化进行多参数和准确功能分析。相关论文于2022年12月5日发表于国际学术期刊《自然-遗传学》杂志。

研究人员研发了成簇规则间隔短回文重复（CRISPR）-Select^TIME、CRISPR-Select^SPACE和CRISPR-Select^STATE，这是一组灵活的敲入方法，可在细胞群中引入遗传变异并跟踪其相对于体内自发变异的绝对频率，其中中性对照突变作为时间，空间或细胞状态可通过流式细胞术测量。在表型上，CRISPR-Select可以确定某种特定变体的致病性，药物反应性/耐药性或促进体内肿瘤发生的概率。

从机制上讲，CRISPR-Select可以通过将变异与运动/侵袭性或任何细胞状态或生化过程与流式细胞术标记物的因果关系联系起来来揭示变异如何引发表型改变。该方法适用于类器官、非转化细胞系或癌细胞系。它准确、定量、快速、简单，可以单孔或96孔高通量形式工作。CRISPR-Select为遗传性疾病的研究、诊断和药物开发提供了一种多样功能变异检测。

据了解，确定与遗传疾病相关数千种基因序列变异（突变）的功能是一项重大挑战。

附：英文原文

Title: Multiparametric and accurate functional analysis of genetic sequence variants using CRISPR-Select

Author: Niu, Yiyuan, Ferreira Azevedo, Catarina A., Li, Xin, Kamali, Elahe, Haagen Nielsen, Ole, Storgaard Srensen, Claus, Frdin, Morten

Issue&Volume: 2022-12-05

Abstract: Determining the functional role of thousands of genetic sequence variants (mutations) associated with genetic diseases is a major challenge. Here we present clustered regularly interspaced short palindromic repeat (CRISPR)-SelectTIME, CRISPR-SelectSPACE and CRISPR-SelectSTATE, a set of flexible knock-in assays that introduce a genetic variant in a cell population and track its absolute frequencies relative to an internal, neutral control mutation as a function of time, space or a cell state measurable by flow cytometry. Phenotypically, CRISPR-Select can thereby determine, for example, pathogenicity, drug responsiveness/resistance or in vivo tumor promotion by a specific variant. Mechanistically, CRISPR-Select can dissect how the variant elicits the phenotype by causally linking the variant to motility/invasiveness or any cell state or biochemical process with a flow cytometry marker. The method is applicable to organoids, nontransformed or cancer cell lines. It is accurate, quantitative, fast and simple and works in single-well or 96-well higher throughput format. CRISPR-Select provides a versatile functional variant assay for research, diagnostics and drug development for genetic disorders.

DOI: 10.1038/s41588-022-01224-7

Source: https://www.nature.com/articles/s41588-022-01224-7

Nature Genetics：《自然—遗传学》，创刊于1992年。隶属于施普林格·自然出版集团，最新IF：41.307
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