Exploring the RNA world, discovering new modes of regulation and how they are used throughout biology from yeast to stem cells.

RNA Tagging

image for RNA TaggingRNA 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

RNA-Tagging Sample Prep 4.3


Lapointe CP, Wilinski D, Saunders HA, Wickens M. 2015 Protein-RNA networks revealed through covalent RNA marks. Nature Methods 12(12):1163-70. PMID:2652440. [PubMed]  [PDF] [Supp. figures]

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