Novel decapping enzyme for cofactor- and “canonically”-capped RNA species

Besides its four canonical nucleotides, RNA molecules can be capped with diverse canonical (N7-methylguanosine (m7G)) and non-canonical chemical modifications (including NAD, FAD, Coenzyme A and Ap(n)A). In ligation-based methods such as 5′ RACE and RNA-seq, decapping of RNAs is useful for detection and identification of RNA containing non-canonical initiating nucleotides. Nudix hydrolases, such as Dcp2 or NudC, are key decapping enzymes that hydrolyze phosphodiester bonds within these caps. Specifically, NudC cleaves the pyrophosphate group in non-canonical caps such as NAD, producing 5′-monophosphorylated RNA suitable for ligation-based assays or targeted degradation. Moreover, NudC hydrolyzes small molecules containing an ADP moiety such as NADH, NAD+, Appp(p)A, ADP-ribose, ADP-glucose and metabolic co-factors such as FAD, Coenzyme A and Acetyl-CoA