Dissecting long non-coding RNAs derived from microRNA genes in hematopoiesis (Illumina short-read sequencing)

Genome-wide transcription produces a complex population of transcripts and a wide variety of functional lncRNAs occasionally evolve from them. Here, we identify and characterize miRNA gene originated lncRNAs (molncRNAs) by employing Pacific Biosciences long-read sequencing. Combining with RNA-sequencing in differentiating hematopoietic erythrocytes, we construct a global molncRNA landscape during human erythropoiesis. Among them, miR-301b and miR-130b originated lncRNA, molnc-301b, orchestrates erythropoiesis independent of miRNAs. Mechanistically, molnc-301b attenuates the transcription of a series of erythropoietic and translation-associated genes by antagonizing SMARCA5's chromatin binding. Furthermore, molnc-301b perturbs translation of key erythroid mRNAs selectively, as GATA1 and FOS. Our findings identify a novel class of lncRNA and reveal the essential role of molnc-301b in erythroid differentiation. Of note, molnc-301b orchestrates erythropoiesis via jointly controlling the transcription and translation of key erythroid regulators. In addition, we finally establish functional molncRNA screen platform by using large-scale pooled CRISPR-based single-cell RNA-seq, and uncover 36 functional molncRNAs in erythrocytes. These results thus unearth a previously unannotated class of lncRNAs and provide a useful platform for the discovery and functional study of molncRNAs. smallRNA-seq, Poly(A)+ RNA-seq, ChIRP-seq, Ribo-seq and RNA-seq datasets.