Single-cell RNAseq of Neurog1-Neurog2-DsRed transduced DRG cells

Cultured adult mouse dorsal root ganglia (DRG) cells exhibit glial sensory progenitor properties in vitro. Therefore, they might be a good starter cell for reprogramming into sensory neurons. Here, we infected them with the retroviral vector Neurog2-Neurog1-DsRed to induce sensory neuron development and analyzed by scRNAseq at 14 days post infection whether infected cells show properties of sensory neurons such as nociceptors. After 10x Genomics, data analysis of 4,549 individual cells indicated the generation of neurons, but at an immature cell state. Cells from dorsal root ganglia culture of adult mice were infected with Neurog2-Neurog1-DsRed, DsRed-positive cells were isolated with Fluorescence-activated cell sorting (FACS) at 14 days post infection and analyzed using scRNAseq.

培养的成年小鼠背根神经节（dorsal root ganglia, DRG）细胞在体外具有胶质细胞感觉祖细胞特性，因此或可作为理想的起始细胞，用于重编程为感觉神经元。本研究使用逆转录病毒载体Neurog2-Neurog1-DsRed感染此类细胞，以诱导感觉神经元分化，并于感染后14天通过单细胞RNA测序（single-cell RNA sequencing, scRNAseq）分析感染细胞是否具备伤害性感受器神经元等感觉神经元的特征。经10x Genomics平台处理后，对4549个单个细胞的数据分析结果显示，成功生成了神经元，但细胞处于未成熟状态。此外，本研究取成年小鼠背根神经节培养细胞，以Neurog2-Neurog1-DsRed进行感染，于感染后14天通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）分离DsRed阳性细胞，并采用单细胞RNA测序完成分析。