7/25/2023 0 Comments Iclip money clip![]() ![]() ![]() These cross-linking-induced mutations in HITS-CLIP and PAR-CLIP can be used as markers to identify the precise RBP binding sites. PAR-CLIP introduces a distinct spectrum of substitutions (T-to-C for 4SU and G-to-A for 6SG). For example, HITS-CLIP utilizes UV cross-linking of proteins with RNA, which introduces either insertions, deletions, or substitutions, depending on the RBPs. This cross-linking process usually introduces mutations in sequence tags at RBP binding sites. PAR-CLIP introduces photoactivatable ribonucleoside analogs, such as 4-thiouridine (4SU) and 6-thioguanosine (6SG), into the RNA of cultured cells to enhance cross-linking efficiency. To increase detection sensitivity, photoactivatable-ribonucleoside-enhanced CLIP (PAR-CLIP) was also developed. For example, high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) was used to identify approximately 30 to 60 nucleotide regions around the peaks of CLIP read clusters that represent binding sites of RNA-binding proteins (RBPs). Recent technological developments, especially the technique of crosslinking immunoprecipitation coupled with high-throughput sequencing (CLIP-seq), have provided powerful tools for studying the roles of RNA regulation in the control of gene expression and the generation of phenotypic complexity. RNA’s diversity in sequence and structure endows it with crucial roles in cell biology. ![]()
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