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The relevance of RNA-protein interactions in regulating the function of mRNA and non-coding RNA is gaining increasing attention. RIP is a powerful technique for studying the physical link between individual proteins and RNA molecules in vivo.
RIP is based on the use of specific antibodies against a protein of interest to pull down RNA binding protein (RBP) and target RNA complexes. Any RNA associated with this protein complex will also be isolated and purified, and RNA bound to the complex can be verified by qPCR or sequencing analysis.
There are several variants of this technology that can be divided into two broad categories: natural and cross-linked RNA immunoprecipitation (live cells are treated by formaldehyde UV light to fix the RNA/protein interactions). Natural RIP can reveal the identity of RNA that binds directly to the protein and its abundance in immunoprecipitated samples, while cross-linked RIP can accurately map the direct and indirect binding sites of the target RBP to RNA molecules.
Living cells are treated with formaldehyde to produce protein-RNA crosslinks between the proximal molecules. Since formaldehyde inactivates cellular enzymes substantially immediately after addition to cells, RIP provides a snapshot of protein-RNA interactions at specific time points and can, therefore, be used for kinetic analysis of events occurring in vivo.
The isolated RNA can be analyzed by several molecular methods, including endpoint RT-PCR and quantitative RT-PCR (if the binding target of RBP is known), microarray or deep sequencing methods.
IntegrateRNA has focused on building efficient systems and procedures to meet the needs and timelines of clients working in RNA research. IntegrateRNA offers the RNA immunoprecipitation services group including expert study designs, immunoprecipitation, and data interpretation, which allows you to characterize the interaction between RNA its bounding protein for functional study. Our RNA immunoprecipitation services will help advance your RNA-related projects and enhance your organization's ability to make sound decisions towards key milestones. If you have any questions don't hesitate to contact us.