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 introduction of RIP-ChIP/Seq
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.
A workflow of RIP-ChIP/Seq
The overall scheme of RIP-ChIP/Seq steps in detail as follows:
Figure 1. Diagram of Basic Protocol of RIP-ChIP
The Merits of RIP-ChIP
Compared with conventional methods and technology, RIP-ChIP/Seq has become increasingly more favored by the researchers because of its own a variety of advantages, including:
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.