RIP chip
The RIP chip ( RNA immunoprecipitation chip , 'RNA immunoprecipitation microarray') is a method of biochemistry for the detection of RNA-protein interactions . The RIP-Chip is a combination of an immunoprecipitation of an RNA-binding protein with an RT-PCR and a microarray and is used to examine the different bound RNA of such a protein.
principle
By immunoprecipitation of a known, mostly recombinant protein, specifically binding RNA sequences can be enriched and purified from an RNA solution . This RNA is then separated from the protein and transcribed into cDNA by RT-PCR. This generated cDNA is used for hybridization with DNA on a microarray.
The RIP-Chip has similarities to the ChIP-on-Chip , which characterizes bound DNA sequences of DNA-binding proteins with a similar principle . The CLIP-Seq , PAR-CLIP and iClip are alternative methods to the RIP chip with DNA sequencing in high-throughput instead of the microarrays.
Individual evidence
- ↑ Townley-Tilson WH, Pendergrass SA, Marzluff WF, Whitfield ML: Genome-wide analysis of mRNAs bound to the histone stem-loop binding protein . In: RNA . 12, No. 10, October 2006, pp. 1853-67. doi : 10.1261 / rna.76006 . PMID 16931877 . PMC 1581977 (free full text).
- ↑ Khalil AM, Guttman M, Huarte M, et al. : Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression . In: Proc. Natl. Acad. Sci. USA . 106, No. 28, July 2009, pp. 11667-72. doi : 10.1073 / pnas.0904715106 . PMID 19571010 . PMC 2704857 (free full text).
- ↑ DG Hendrickson, DJ Hogan, D. Herschlag, JE Ferrell, PO Brown: Systematic identification of mRNAs recruited to argonaute 2 by specific microRNAs and corresponding changes in transcript abundance. In: PloS one. Volume 3, number 5, 2008, ISSN 1932-6203 , p. E2126, doi : 10.1371 / journal.pone.0002126 . PMID 18461144 , PMC 2330160 (free full text).