As former bodybuilder turned California Governator, Arnold Schwarzenegger, once said, “Great abs are made in the kitchen…” That may be, but when it comes to epigenetics research, great abs are made by great labs and suppliers, and they’re put to work in immunoprecipitation applications like RIP Chip.
Recently, Anke van den Berg and colleagues at the University of Groningen in the Netherlands further streamlined the RIP-Chip workout, enabling broad coverage of all miRNA targets while also providing the option to refine the assay for more targeted applications. The workflow goes like so:
- Ribonucleoprotein IP (RIP) against Argonaute 2 to select for miRNAs and their targets
- Subsequent microarray analysis to identify targets
- To refine experiment, dose cells with anti-miR-YFM (Your Favorite MicroRNA)
- mRNA lost in subsequent pull down are considered targets of YFM
Applying the RIP-Chip workout allowed the Dutch team to identify miRNA targets in a Hodgkin Lymphoma cell background, where they enriched 10 to 30% of all transcripts as the miRNA “targetome”. Their subsequent refining work with anti-miR-17/20/93/106 identified 1,189 gene transcripts thought to be regulated by these miRNAs. The team managed to validate 8 of 10 of these experimentally.