RNA结合蛋白与RNA互作鉴定技术

RNA结合蛋白（RNA binding proteins，RBPs）通过与RNA相互作用，广泛参与到RNA的剪切、转运、编辑、胞内定位及翻译调控等过程中。RNA领域尤其是非编码RNA（non-coding RNA，ncRNA）研究的快速发展，催生了多种RBPs RNAs相互作用鉴定技术。这些技术反之又推动了 RNA领域的研究进程。本文对紫外交联免疫沉淀（ultraviolet crosslinking and immunoprecipitation，CLIP）,CLIP cDNA文库高通量测序 (high-throughput sequencing of CLIP cDNA library，HITS-CLIP),光活化核苷增强的CLIP(photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation，PAR-CLIP),单核苷酸分离CLIP (individual nucleotide resolution CLIP，iCLIP),TRIBE (targets of RNA-binding protein identified by editing),RNA 标记，相互作用组捕获(interactome capture，IC) 和SerIC (serial RNA interactome capture)等RBPs-RNAs相互作用鉴定技术的基本原理和优缺点以及应用进行综述。

Techniques to Identify the Interactome of RNA Binding Proteins and RNAs

RNA binding proteins (RBPs) are widely involved in RNA related processes such as splicing, transport, editing, cellular localization, and translational regulatory via interacting with target RNAs. With the rapid development of RNA research, especially in non-coding RNA (ncRNA), a variety of techniques of RBPs-RNAs interaction identification were created, which in turn contributed to the understanding of molecular mechanisms of RNAs processing and function. Herein, we review the basic principles, advantages and disadvantages, and applications of some important techniques that are mainly used for studying the interactome of RBPs-RNAs, including CLIP (ultraviolet crosslinking and immunoprecipitation), HITS-CLIP (high-throughput sequencing of CLIP) analysis of cDNA library, PAR-CLIP (photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation), iCLIP (individual nucleotide resolution CLIP), TRIBE (targets of RNA-binding protein identified by editing), RNA tagging, IC (interactome capture) and SerIC (serial RNA interactome capture).