Gene expression and 4sUDRB-seq for NF90/NF110 of human scramble and KD HeLa cells.

We have optimized the metabolic tagging of newly transcribed RNAs by 4-thiouridine (4sU) to identify newly-transcribed (nascent) circRNAs from human HeLa cells. Overall design: Poly(A)+ RNA-seq for gene expression, ribo–/poly(A)– for steady-state circRNA, Ribo– 4sUDRB-seq for characterizing nacent circRNA biogenesis and iCLIP-seq of NF90 for RNA binding analysis.