Accurate quantification by sequencing (AQ-seq) of small RNA species co-immunoprecipitated with human C12orf29 from HEK293A cell lines

ATP-grasp superfamily enzymes contain a hand-like ATP-binding fold and catalyze a variety of reactions using a similar catalytic mechanism. More than 30 protein families are categorized in this superfamily, and they are involved in a plethora of cellular processes and human diseases. We identify C12orf29 as an atypical ATP-grasp enzyme that ligates RNA. C12orf29 E250A binds small RNA species in cells, which are potential substrates of C12orf29. To identify these species, we perform Flag-immunoprecipitation (IP) of C12orf29 WT or E250A from HEK293A cell lines, extracted RNA and purified small RNA species by Urea-PAGE to perform Accurate quantification by sequencing (AQ-seq)