CLIP相关技术的研究进展

Cross-linking and immunoprecipitation assay and its development

在真核生物基因表达的转录后调节中,RNA结合蛋白(RBP)起着关键作用,很多RBP的异常与人类疾病的发生密切相关。自2000年的RNA免疫沉淀和芯片分析方法(RNA immunoprecipitation with differential display or microarray analysis,RIP-ChIP)出现以来,人们开始就RBP与RNA相互作用进行了系统而广泛的研究。经过改良和发展,基于体内实时紫外交联免疫沉淀法(ultraviolet crosslinking and immunoprecipitation,CLIP)、交联免疫沉淀cDNA文库高通量测序法(high-throughput sequencing of CLIP cDNA library,HITS-CLIP)、光催化核糖核苷增强交联和免疫沉淀法(photoactivatable-ribonucleoside-enhanced crosslinking and immunprecipitation,PAR-CLIP)以及提高个别核苷酸分辨率交联和免疫共沉淀法(individual nucleotide resolution CLIP,iCLIP)等RIP-ChIP衍生方法相继产生,使用这些方法,可以解析RBP的RNA识别特异性,而且通过与高通量测序技术结合,可以实现转录组尺度的RBP的靶序列的鉴定,分辨率也得到极大提高。该文就RNA与蛋白的相互作用的基本原理及其研究进展、相关技术存在的问题以及发展趋势进行简要综述。

It′s reported that RNA-binding proteins （ RBP） play key roles in post-transcriptional regulation of eukaryotic genes.The aberrations of RBP are associated with a large number of human disorders , particularly autoimmune and neuro-logic diseases .The interaction between RNA and proteins has been widely explored since the development of the method known as RNA immunoprecipitation with differential display or microarray analysis （RIP-ChIP） around the year of 2000. Since then, diverse derivatives of the RIP-ChIP, such as ultraviolet crosslinking and immunoprecipitation （ CLIP）, high-throughput sequencing of CLIP cDNA library （HITS-CLIP）, photoactivatable -ribonucleoside-enhanced crosslinking and immunoprecipitation （ PAR-CLIP） , and individual nucleotide resolution CLIP （ iCLIP ） have been developed .All these methods have some advantages over the original RIP-ChIP and greatly facilitate the study of RBP-RNA interactions .Addi-tionally , aided by the next-generation sequencing , transcriptome-wide identification of RBP target sites has become possible and the RNA-binding site resolution of RBP has also improved to some degree .We introduced the basic principles and processes of the interactions between proteins and RNA , focusing on the advantages , disadvantages and prospect of the present genome-wide version of CLIP .