Complete male-to-female sex reversal in mice lacking the miR-17~92 cluster [RNA-seq]

The deletion of a single miRNA cluster, miR-17~92, is sufficient to induce primary sex reversal in XY mice. The expression of the testis determining gene, Sry, is delayed in embryonic XY miR-17~92 knockout gonads, which immediately activate the ovarian genetic program. Single cell RNA-seq analysis shows that Sertoli cell differentiation is highly reduced, delayed and unable to trigger testis differentiation. Consistent with the well-known role of miRNAs in gene regulation, the expression of target genes of miR-17~92 is not stabilized in XY mutant gonads at E11.5, affecting, in turn, the fine regulation of large gene networks involved in mammalian sex determination. Our results reveal that the miR-17~92 cluster is a novel sex-determining factor that modulates several gene networks required for accurate timing of Sry expression and Sertoli cell differentiation during testis determination and early differentiation. bulk RNA-seq, single cell RNA-seq as well as IF and other characterizations of a transgenic mice for miR-17~92 cluster Alicia Hurtado单个微RNA（microRNA, miRNA）簇miR-17~92的缺失即可诱导XY小鼠发生原发性性反转。胚胎期XY型miR-17~92敲除性腺中，睾丸决定基因Sry（Sex-determining Region Y）的表达出现延迟，且该性腺会立即激活卵巢遗传程序。单细胞RNA测序（single cell RNA-seq）分析显示，支持细胞（Sertoli cell）分化程度显著降低、进程延迟，且无法启动睾丸分化。与微RNA在基因调控中的经典作用一致，miR-17~92的靶基因在胚胎发育第11.5天（E11.5）的XY突变性腺中未得到稳定表达，进而干扰了哺乳动物性别决定相关大型基因网络的精细调控。本研究结果表明，miR-17~92簇是一种新型性别决定因子，可调控睾丸决定与早期分化过程中Sry表达时序及支持细胞分化所需的多个基因网络。本数据集包含miR-17~92簇转基因小鼠的批量RNA测序（bulk RNA-seq）、单细胞RNA测序（single cell RNA-seq）以及免疫荧光（Immunofluorescence, IF）等多种表征数据，研究者为Alicia Hurtado。