Fig. 3.

Process diagrams of representative nascent RNA-seq methods. a Schematic of GRO-seq. In this approach, nascent RNAs are labeled with 5Br-UTP and immunoprecipitated with the antibody anti-Br-UTP; the isolated RNAs subsequently undergoes deep sequencing. b Schematic of methods based on base mutation for nascent RNA detection. Nascent RNAs are labeled with a thiol-labeled nucleoside (s⁴U or s⁶G), and these newly synthesized RNAs can then be isolated and treated with specific chemical reagents, such as thiol (SLAM-seq) and acrylonitrile (AMUC-seq), leading to a change in the base-pairing manner of metabolically incorporated nucleosides