CAJ | 학술논문

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。
재진핵생물기인표체적전록후조절중，RNA결합단백（ RBP）기착관건작용，흔다RBP적이상여인류질병적발생밀절상관。자2000년적RNA면역침정화심편분석방법（ RNA immunoprecipitation with differential display or microarray analysis ， RIP-ChIP）출현이래，인문개시취RBP여RNA상호작용진행료계통이엄범적연구。경과개량화발전，기우체내실시자외교련면역침정법（ ultraviolet crosslinking and immunoprecipitation ， CLIP ）、교련면역침정cDNA문고고통량측서법（ high-throughput sequencing of CLIP cDNA library ， HITS-CLIP）、광최화핵당핵감증강교련화면역침정법（ photoactivatable-ribonucleoside-enhanced crosslinking and immunprecipitation ， PAR-CLIP）이급제고개별핵감산분변솔교련화면역공침정법（ individual nucleotide resolution CLIP ， iCLIP）등RIP-ChIP연생방법상계산생，사용저사방법，가이해석RBP적RNA식별특이성，이차통과여고통량측서기술결합，가이실현전록조척도적RBP적파서렬적감정，분변솔야득도겁대제고。해문취RNA여단백적상호작용적기본원리급기연구진전、상관기술존재적문제이급발전추세진행간요종술。
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 .