TRAP-TAGGING: A NOVEL METHOD FOR THE IDENTIFICATION AND PURIFICATION OF RNA-PROTEIN COMPLEXES

摘要

With the recent completion of several genome sequencing projects, scientists have embarked on comprehensive attempts to unravel all of the interactions amongst their gene products. While many proteomics efforts are well under way, little attention has been paid t o the RNA products of these genes. The variety and scope of roles that RNA molecules play in biological processes is only now beginning to be appreciated. Consequently, "ribonomics" will likely be the next focus of genomic efforts. Unfortunately, conventional methods for the isolation an d identification of specific RNA-protein complexes are plagued by a number of problems not encountered in genomics or proteomics. Here we describe a method that circumvents these problems. The TRAP (Tandem RNA Affinity Purification) tag is a dual RNA tagging system tha t facilitates gentle purification of RNA molecules along with the proteins, RNAs and other small molecules specifically associated with them.