Cross-linking and immunoprecipitation assay and its development / 军事医学

ABSTRACT

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 .