RIP-Seq Service
In eukaryotic organisms, a tight regulation of gene expression is required for complex processes such as development, differentiation, cell specification, and response of each cell to environmental signals. Post-transcriptional regulation as an essential step of gene expression involves the interactions of RBPs, mRNA and noncoding RNAs (e.g., microRNAs) that affect splicing, nuclear export, subcellular localization, mRNA decay and translation. Thus, more and more researchers attempt to explore this complex regulation mechanism through the identification of the RNAs associated with RBPs.
However, traditional research methods and technologies are limited in their own features. RIP-ChIP/Seq technique as a powerful in vivo, high-throughput technique for identifying specific associated RBP targets from cell extracts employs immunoprecipitation of endogenously formed mRNP complexes using an antibody specific to the RBP, followed by purification of associated RNAs and their global, quantitative analysis using a genomic read-out such as microarrays, RT-qPCR analysis, or next generation sequencing analysis. RIP-ChIP/Seq provides the possibility to better understand the mechanism of post-transcriptional regulation and even gene expression in vivo.
As an expert in the field of studying protein-nucleic acid interactions, Profacgen provides a powerful technique, RNA-binding proteins immunoprecipitation microarray or next generation sequencing analysis (RIP-ChIP/Seq), which has been widely used for isolating and identifying mRNA and noncoding RNAs associated with RNA-binding proteins (RBPs) in vivo.
The overall scheme of RIP-ChIP/Seq steps in detail as follows:
Services
As an expert in protein-nucleic acid interaction research, Profacgen recruited biochemical experts and omics experts and jointly established the efficient RIP-seq technology platform. We provide one-stop service from RNA sampling to bioinformatics analysis, and strictly verify each step including sample quality control, library quality control, and sequencing data quality control to ensure the accuracy and reliability of sequencing data. Our sequencing platform is based on Illumina/Solexa and optimized to fully extract useful information (i.e., the original RNA sequence combined by protein) from the short sequence and obtain effective information of RNA-protein interactions.
Our experimental procedure is as follows:
- Cell harvesting
- Isolation of nuclei and lyse of nuclear pellets
- Chromatin shearing
- RNA immunoprecipitation
- Wash off unbound material
- Purification of RNA that is bound to immunoprecipitated RBP
- Reverse transcription of RNA to cDNA and analysis by qPCR, microarray, or sequencing
Significantly, based on the customer requirements, Profacgen could also provide personalized testing services on protein-nucleic acid interactions, like CLIP-seq, CHIP, ChIRP-seq services, to further provide strong support for studying the complex regulation mechanism of gene expression.
Profacgen has accumulated lots of experience in nucleic acid-protein interaction research. Our professional technical team can provide customers with efficient RIP-seq and many related featured services. Our competitive prices and extensive expertise have earned us the trust of our collaborators. Contact us to find out how Profacgen could be of assistance.
References
- Moore, K.S.; Hoen, P.A. Computational approaches for the analysis of RNA-protein interactions: A primer for biologists. Journal of Biological Chemistry. 2019.
- Selth, L.A.; et al. RNA immunoprecipitation to determine RNA-protein associations in vivo. Cold Spring Harbor Protocols. 2009.