Visualizing RNA Cytidine Acetyltransferase Activity by Northern Blotting

Cytidine acetyltransferases are an emerging class of nucleic-acid-modifying enzymes responsible for the establishment of N⁴ -acetylcytidine (ac4C) in RNA. In contrast to histone acetyltransferases, whose activity is commonly studied by western blotting, relatively few methods exist for quickly assessing the activity of cytidine acetyltransferases from a biological sample of interest or the distribution of ac4C across different RNA species. In this protocol, we describe a method for analysis of cellular cytidine acetyltransferase activity using dot- and immuno-northern-blotting-based detection. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Detection of N⁴ -Acetylcytidine in RNA by dot blotting Basic Protocol 2: Visualizing N⁴ -Acetylcytidine Distribution in RNA by northern blotting.