Single-cell transcriptome analysis of mouse main olfactory epithelium

The goal of scRNA-seq is to explore whether olfactory sensory neurons (OSNs) that display a specific olfactory receptor (OR) express a distinct transcriptional program from OSNs that display a different OR in a large and unbias scale. Olfactory epithelial single cell suspensions were made using 3 male and 3 female mice. Single-cell sequencing libraries were prepared using Chromium Single Cell 3' Reagent Kit V3 (10X Genomics) according to the manufacturer's guidelines. Libraries were sequenced using 150 cycles of paired end reads on Illumina Hiseq4000 and Novaseq6000 instruments (Novogene). The sequencing reads were processed using the DolphinNext Single Cell-10X Genomics pipeline (https://dolphinnext.umassmed.edu/index.php?np=1&id=420, default settings except STAR v2.6.1 was used for alignment). Overall design: Olfactory epithelial single cell suspensions made from 3 male and 3 female adult mice. Single-cell sequencing libraries were prepared using Chromium Single Cell 3' Reagent Kit V3 (10X Genomics) according to the manufacturer's guidelines. Libraries were sequenced using 150 cycles of paired end reads on Illumina Hiseq4000 and Novaseq6000 instruments (Novogene)

单细胞RNA测序（scRNA-seq）的核心目标在于，以大规模无偏的研究尺度，探究表达特定嗅觉受体（olfactory receptor, OR）的嗅觉感觉神经元（olfactory sensory neurons, OSNs）是否与表达其他嗅觉受体的嗅觉感觉神经元存在差异化转录程序。本实验采用3只雄性与3只雌性小鼠制备嗅觉上皮单细胞悬液。依照制造商操作指南，使用10X Genomics公司的Chromium单细胞3'端试剂试剂盒V3构建单细胞测序文库。文库于Illumina Hiseq4000与Novaseq6000测序仪上采用150轮双端读长进行测序，测序服务由诺禾致源（Novogene）提供。测序读段通过DolphinNext单细胞-10X Genomics分析流程（默认参数，仅比对步骤采用STAR v2.6.1版本，流程链接：https://dolphinnext.umassmed.edu/index.php?np=1&id=420）进行处理。整体实验设计：采用3只雄性与3只雌性成年小鼠制备嗅觉上皮单细胞悬液；依照制造商操作指南，使用10X Genomics公司的Chromium单细胞3'端试剂试剂盒V3构建单细胞测序文库；文库于Illumina Hiseq4000与Novaseq6000测序仪上采用150轮双端读长进行测序，测序服务由诺禾致源（Novogene）提供。