Next Generation Sequencing Facilitates Quantitative Analysis of chromatin interaction of lncRNA Gm4665 with regulatory region of target genes in male germ cells.. Next Generation Sequencing Facilitates Quantitative Analysis of chromatin interaction of lncRNA Gm4665 with regulatory region of target genes in male germ cells.

Purpose: The goals of this study are to examine the possibility that Gm4665 binds to regulatory region of target genes loci in male germ cells. Methods: we performed the Chromatin Isolation by RNA Purification (ChIRP) assay to examine the possibility that Gm4665 binds to regulatory region of target genes. Germ cell isolated from the testis of wild-type mouse were cross-linked, lysed and sonicated, and then incubated with probesets of Gm4665. Following hybridization of biotinylated probesets to Gm4665 transcript, Gm4665-bound chromatin was retrieved. The purified DNA fragments were sequenced using high throughput next generation sequencing (Illumina HiseqXten-PE150) Results: Analysis of the retrieved chromatin fragments by deep sequencing (ChIRP-seq) revealed Gm4665 occupancy sites were genome wide, specifically abundant enrich in intronic regions. Additionally, nearly 30% of chromatin fragments were enriched in the promoter regions of gene loci Overall design: Examination of chromatin interaction of lncRNA Gm4665 with regulatory region of target genes in male germ cells