Small Nucleolar RNAs in the Dlk1-Gtl2 Imprinting Locus Maintains LT-HSC Self-renewal by Regulation of rRNA Modification and Translation [scRNA-Seq]. Small Nucleolar RNAs in the Dlk1-Gtl2 Imprinting Locus Maintains LT-HSC Self-renewal by Regulation of rRNA Modification and Translation [scRNA-Seq]

Small nucleolar RNA (snoRNA) are non-coding RNAs, which participate in the cleavage and chemical modification of ribosomal RNAs (rRNAs) and small nuclear RNAs. However, the roles of snoRNA in homeostasis of hematopoietic stem cells(HSCs) have not been studied. We isolated four hematopoietic stem and progenitor cells (HSPCs) (LT-HSCs, IT-HSCs, ST-HSCs, and MPPs) from the bone marrow (BM) of C57BL/6 mice and performed small RNA-seq. We found SnoRNAs belonging to SNORD113-114 cluster were specifically enriched in LT-HSCs, and their expression decreased dramatically with HSC differentiation. To explore function of snoRNAs in SNORD113-114 cluster in HSCs, we established maternally KO mice by CRISPR-Cas9 technology. Here, we used scRNA-seq to examine HSC RNA profiles influenced by SNORD113-114 cluster KO. Overall design: Single cell RNA profiles of HSC from WT/Mat KO mice.

核仁小RNA（small nucleolar RNA, snoRNA）是一类非编码RNA，参与核糖体RNA（ribosomal RNAs, rRNAs）与小核RNA的剪切及化学修饰过程。目前学界尚未阐明snoRNA在造血干细胞（hematopoietic stem cells, HSCs）稳态调控中的作用。本研究从C57BL/6小鼠骨髓（bone marrow, BM）中分离得到四类造血干祖细胞（hematopoietic stem and progenitor cells, HSPCs），即LT-HSCs、IT-HSCs、ST-HSCs及MPPs，并开展了小RNA测序（small RNA-seq）。结果显示，隶属于SNORD113-114基因簇的snoRNA在LT-HSCs中特异性富集，且其表达水平随HSC分化进程显著下调。为探究该基因簇snoRNA在HSCs中的功能，本研究通过CRISPR-Cas9技术构建了母系敲除小鼠模型。随后，本研究利用单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）分析了SNORD113-114基因簇敲除对HSC转录组谱的影响。整体实验设计：对野生型（wild-type, WT）与母系敲除（maternally knockout, Mat KO）小鼠的造血干细胞进行单细胞RNA谱分析。