Identification of Immune Modulatory miRNAs by miRNA Enrichment via RNA Affinity Purification.

Abstract

Immune escape by cancer cells can be triggered by aberrant expression of immunological key players, which can be achieved by distinct molecular mechanisms including immune modulatory miRNAs. One suitable method to identify miRNAs that specifically target immune relevant molecules is the miRNA enrichment via RNA affinity purification method named miTRAP (miRNA trapping by RNA in vitro affinity purification). Here, we present a detailed protocol for construct preparation, RNA immobilization via MS2BP-MBP to beads, miRNA enrichment, and elution followed by analysis of the obtained miRNA candidates via qRT-PCR.