RAP Service

RAP (RNA Antisense Purification) captures the target RNA of its hybridization by biotin-labeled antisense oligonucleotide probes, which can be purified to obtain endogenous RNA and the proteins and nucleic acids bound to it. Combining RAP with high-throughput sequencing (RAP-Seq) and mass spectrometry (RAP-MS) technologies allows the study of RNA-chromatin interactions, the identification of proteins that interact with RNA, and the study of transcriptional regulation between multiple RNAs at the transcriptome level. RAP has been able to fully map RNA-DNA interactions in vivo.

To study the mechanism of action of RNA molecules, such as lncRNAs, and to identify proteins that interact directly and specifically with target RNA molecules, Creative Proteomcis presents a holistic research solution for RAP technology that can reveal the study of RNA-DNA, RNA-protein, and RNA-RNA interactions.

What We Can Provide

• RAP-MS: Biotin-labeled target RNA was synthesized in vitro (Control group used scramble RNA). Subsequently, it was added to non-denatured lysed biotin samples and incubated to replace the endogenous RNA. The biotin is pull-down by immobilized avidin, and the resulting RBP is identified and quantified by LC-MS/MS, from which the interacting protein profile of the target RNA is constructed.
• RAP-Seq

Service Advantages

To robustly capture lncRNA, RAP uses probes that are 120-nt long probes. Long probes have the advantage of reduced background noise. the RAP technique does not require prior knowledge of the lncRNA binding domains involved in chromatin interactions. Tiling oligonucleotides over the entire target RNA allows all potential hybridization sites to be fully exploited to ensure capture of the target RNA and its bound proteins and DNA even in the presence of extensive protein- RNA interactions, RNA secondary structure or partial RNA degradation.

Sample Requirements

Live cells: ≥ 10⁷

Fresh tissue: ≥ 0.5g

RNA information, species information

Delivery of Results

• Assay report (experimental instruments, reagents, methods, results, conclusions)
• RAP probe set sequence
• RAP-MS: peptide fingerprinting, library search results
• RAP-RNA: downstream target gene name, PCR electrophoresis map, or high-throughput sequencing results