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Urb-RIP – An Adaptable and Efficient Approach for Immunoprecipitation of RNAs and Associated RNAs/Proteins
Abstract
<div><p>Post-transcriptional regulation of gene expression is an important process that is mediated by interactions between mRNAs and RNA binding proteins (RBP), non-coding RNAs (ncRNA) or ribonucleoproteins (RNP). Key to the study of post-transcriptional regulation of mRNAs and the function of ncRNAs such as long non-coding RNAs (lncRNAs) is an understanding of what factors are interacting with these transcripts. While several techniques exist for the enrichment of a transcript whether it is an mRNA or an ncRNA, many of these techniques are cumbersome or limited in their application. Here we present a novel method for the immunoprecipitation of mRNAs and ncRNAs, Urb—RNA immunoprecipitation (Urb-RIP). This method employs the RRM1 domain of the “resurrected” snRNA-binding protein Urb to enrich messages containing a stem-loop tag. Unlike techniques which employ the MS2 protein, which require large repeats of the MS2 binding element, Urb-RIP requires only one stem-loop. This method routinely provides over ~100-fold enrichment of tagged messages. Using this technique we have shown enrichment of tagged mRNAs and lncRNAs as well as miRNAs and RNA-binding proteins bound to those messages. We have confirmed, using Urb-RIP, interaction between RNA PolIII transcribed lncRNA BC200 and polyA binding protein.</p></div- Dataset
- Dataset
- Biophysics
- Biochemistry
- Microbiology
- Cell Biology
- Genetics
- Molecular Biology
- Immunology
- Developmental Biology
- Mental Health
- Infectious Diseases
- Biological Sciences not elsewhere classified
- Physical Sciences not elsewhere classified
- RNA-binding proteins
- MS 2 protein
- stem-loop tag
- technique
- lncRNA BC 200
- RNA binding proteins
- gene expression
- RRM 1 domain
- RBP
- mRNA
- RNP
- Efficient Approach
- non-coding RNAs
- Urb-RIP
- post-transcriptional regulation
- enrichment
- polyA binding protein
- RNA PolIII
- novel method
- MS 2 binding element