Profiling the lncRNA interqaction network in the Oct4 promoter of fibroblasts in reprogramming by chromatin in situ reverse transcription trap sequencing

“chromatin-lncRNA in situ reverse transcription trap sequencing” (CRIST-Seq) approach was used to profile the lncRNA interactome in the promoter of stem cell core factors Oct4 Fibroblasts (FIB) were transfected with CRISPR Cas9 and two gRNAs derived from the gene promoter. Cells were crosslinked with formaldehyde to fix the chromatin structure. Long noncoding RNAs (lncRNAs) in the chromatin complex of the OCT4 promoter was in situ reverse transcribed with biotin-dCTP. After immunoprecipitation with Cas9 antibody and streptavidin bead purification, lncRNA derived cDNAs were used for Illumina library sequencing.