RNA Tagging is a simplified and unambiguous approach to analyze protein-RNA interactions in live cells, tissues, or whole animals. In RNA Tagging, we express an RNA-binding protein of interest fused to a C. elegans poly(U) polymerase, PUP-2. The chimera “tags” the RNAs it binds in vivo with 3’ terminal uridines. Tagged RNAs are then identified from a pool of total RNA using both targeted and high-throughput assays. The strategy is independent of crosslinking and protein purification steps.
Below we provide pertinent information to the approach and the reagents we have developed. Please contact us if you have questions, comments, concerns, or improvements.
Current Lab Protocol
Plasmids and Sequences
RNA Tagging Plasmid List
RT4_pBSII_PUF3-3’flank_PUP2_3HA_URA3_PUF3 5′ flank
RT5_pBSII_PUF3-3’flank_PUP2-DADA_3HA_URA3_PUF3 5′ flank