RtcB RNAseq to map cyclic phosphate-bearing RNA fragments during poly I:C treatment

RNase L is activated by 2-5A, which is synthesized specifically upon dsRNA sensing. Activated RNase L cleaves cellular RNAs, generating fragments that bear 2'-3' cylic phosphate and/or 5'-OH. Here we perform cyclic-phosphate capture and RNA-seq to map the precise sites at which cleavage is induced during dsRNA stress. Overall design: A549 cells were treated +/- poly I:C for one hour and total RNA was extracted. Custom library prep was performed to capture and enrich fragments with a 2'-3' cyclic phosphate using RtcB enzyme, followed by RNA-seq.