Identification of N⁶-methyladenosine reader proteins.

The reversible N⁶-methyladenosine (m⁶A) modification of eukaryotic messenger RNAs (mRNAs) is a widespread regulatory mechanism that impacts every step in the mRNA life cycle. The effect of m⁶A on mRNA fate depends on the binding of "m⁶A reader" proteins - RNA binding proteins that specifically bind to RNAs containing m⁶A. Here, we describe an RNA pull-down method that can be used to identify novel m⁶A reader proteins starting from a known m⁶A-modified site in cellular or viral RNA. We further describe how a combination of immunoprecipitation-based sequencing methods can be used to identify m⁶A-modified sites bound by an m⁶A reader protein on a transcriptome-wide level. The discovery of new m⁶A reader proteins and their m⁶A-modified targets would provide further insight into the mechanisms and functions of m⁶A in the cell.

Duke Scholars

Author Katherine Zhou