Genome-wide CRISPR-Cas9 Interrogation of Splicing Networks Reveals a Mechanism for Recognition of Autism-Misregulated Neuronal Microexons
Genome-wide CRISPR-Cas9 Interrogation of Splicing Networks Reveals a Mechanism for Recognition of Autism-Misregulated Neuronal Microexons
复制标题
DOI：
10.1016/j.molcel.2018.10.008
复制
发表时间：
2018-11-01
期刊：
影响因子：
16
通讯作者：
Blencowe, Benjamin J.
中科院分区：
文献类型：
--
作者：
Gonatopoulos-Pournatzis, Thomas;Wu, Mingkun;Blencowe, Benjamin J.
可变剪接对于多种细胞、发育和病理过程至关重要。然而，控制单个剪接事件的完整因子网络尚不清楚。在此，我们描述了一种基于CRISPR的策略，用于在全基因组范围内阐明控制剪接的途径，并将其应用于在神经系统发育中具有重要功能且在自闭症中通常失调的微外显子。大约200个与功能多样的调控层相关且富含与自闭症的遗传关联的基因控制着神经元微外显子。值得注意的是，广泛表达的RNA结合蛋白Srsf11和Rnps1直接、优先且频繁地共同激活这些微外显子。这些因子与神经元剪接调节因子Srrm4以及一个二分内含子剪接增强子元件形成关键相互作用，以促进剪接体的形成。因此，我们的研究提供了一个用于识别整个剪接调控途径的通用系统，并进一步揭示了在自闭症中被破坏的神经元微外显子定义的一种常见机制。
Alternative splicing is crucial for diverse cellular, developmental, and pathological processes. However, the full networks of factors that control individual splicing events are not known. Here, we describe a CRISPR-based strategy for the genome-wide elucidation of pathways that control splicing and apply it to microexons with important functions in nervous system development and that are commonly mis-regulated in autism. Approximately 200 genes associated with functionally diverse regulatory layers and enriched in genetic links to autism control neuronal microexons. Remarkably, the widely expressed RNA binding proteins Srsf11 and Rnps1 directly, preferentially, and frequently co-activate these microexons. These factors form critical interactions with the neuronal splicing regulator Srrm4 and a bi-partite intronic splicing enhancer element to promote spliceosome formation. Our study thus presents a versatile system for the identification of entire splicing regulatory pathways and further reveals a common mechanism for the definition of neuronal microexons that is disrupted in autism.