Romano Giulia, Klima Raffaella, Feiguin Fabian
International Centre for Genetic Engineering and Biotechnology, Trieste Italy, Padriciano 99, 34149 TS, Italy.
Department of Life and Environmental Sciences, University of Cagliari, 09042 Monserrato, Cagliari, Italy.
Bio Protoc. 2021 Dec 5;11(23):e4250. doi: 10.21769/BioProtoc.4250.
To determine the molecular and functional interactions between RNA-binding proteins (RBPs) and their targets RNAs, is of fundamental importance to understand the dynamic organization of the nervous system in health and disease. Nevertheless, this task has remained elusive due to the lack of specific protocols and experimental systems that would allow the combination of biochemical analysis with functional genetics. In this manuscript, we describe a trustworthy and detailed methodology to establish the molecular organization and intracellular function of RBPs/RNA multimeric complexes in a cell type-defined manner by using the powerful GAL4/UAS system for gene expression in . Graphic abstract: Immunoprecipitation for protein-RNA interaction in .
确定RNA结合蛋白(RBPs)与其靶RNA之间的分子和功能相互作用,对于理解健康和疾病状态下神经系统的动态组织至关重要。然而,由于缺乏能够将生化分析与功能遗传学相结合的特定方案和实验系统,这项任务仍然难以捉摸。在本手稿中,我们描述了一种可靠且详细的方法,通过使用强大的GAL4/UAS系统在细胞类型定义的方式下建立RBPs/RNA多聚体复合物的分子组织和细胞内功能。图形摘要:用于蛋白质-RNA相互作用的免疫沉淀。