小柯机器人

德国慕尼黑技术大学医学院Roland Rad、Juan J. Montero团队，利用CasRx对长非编码RNA（lncRNA）的依赖性进行了基因组规模的泛癌症检测。这一研究成果于2024年2月26日发表在国际学术期刊《自然-方法学》上。

研究人员利用Cas13d/CasRx研发了基因组规模的lncRNA转录组筛选。研究发现，RNA靶向克服了其他筛选方法的固有限制，从而大大扩展了lncRNA组的可探索空间。它支持分子和表型数据整合以确定筛选结果的背景或推断lncRNA的功能，研究将该筛选系统应用于泛癌症研究。因此，针对包含24,171个lncRNA基因，研究人员建立了一个称为Albarossa的多重gRNA文库，从而解决了转录组规模和组织特异性所带来的挑战。

其合理的设计结合基于表达、进化保护和组织特异性进行目标优先排序，从而兼顾了高发现力和泛癌症代表性以及可扩展的实验通量。在跨癌症中类的应用中，该筛选平台发现了许多具有特异性和共性的lncRNA。该工作为系统探索健康和疾病中lncRNA的生物学功能奠定了基础。

据了解，尽管lncRNA在转录组中占主导地位，但它们的功能在很大程度上是未知的。lncRNA与编码序列和调控序列的广泛重叠限制了通过DNA定向扰动对其进行系统研究。

附：英文原文

Title: Genome-scale pan-cancer interrogation of lncRNA dependencies using CasRx

Author: Montero, Juan J., Trozzo, Riccardo, Sugden, Maya, llinger, Rupert, Belka, Alexander, Zhigalova, Ekaterina, Waetzig, Paul, Engleitner, Thomas, Schmidt-Supprian, Marc, Saur, Dieter, Rad, Roland

Issue&Volume: 2024-02-26

Abstract: Although long noncoding RNAs (lncRNAs) dominate the transcriptome, their functions are largely unexplored. The extensive overlap of lncRNAs with coding and regulatory sequences restricts their systematic interrogation by DNA-directed perturbation. Here we developed genome-scale lncRNA transcriptome screening using Cas13d/CasRx. We show that RNA targeting overcomes limitations inherent to other screening methods, thereby considerably expanding the explorable space of the lncRNAome. By evolving the screening system toward pan-cancer applicability, it supports molecular and phenotypic data integration to contextualize screening hits or infer lncRNA function. We thereby addressed challenges posed by the enormous transcriptome size and tissue specificity through a size-reduced multiplexed gRNA library termed Albarossa, targeting 24,171 lncRNA genes. Its rational design incorporates target prioritization based on expression, evolutionary conservation and tissue specificity, thereby reconciling high discovery power and pan-cancer representation with scalable experimental throughput. Applied across entities, the screening platform identified numerous context-specific and common essential lncRNAs. Our work sets the stage for systematic exploration of lncRNA biology in health and disease.

DOI: 10.1038/s41592-024-02190-0

Source: https://www.nature.com/articles/s41592-024-02190-0

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
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