RNA-seq of the lacrimal gland epithelial tissue in control vs Pea3-depleted tissue in mouse embyro

The goal of the study is to find the transcriptional targets downstream of Pea3-mediated signaling in the lacrimal gland epithelium. In order to look for the potential targets, we performed laser capture microdissection of the lacrimal gland epithelial tissue at embryonic day E14.5 from control embryos as well as mutant embryos containing epithelium-specific deletion of Pea3. After tissue harvest, RNA was extracted. Conversion to cDNA as well as amplification was performed by Clontech SMART-seq v4 Ultra low input RNA kit, as well as cDNA library construction was performed using Nextera XT DNA library preparation kit by core facility at Columbia university prior to RNA sequencing. After preparation of cDNA library, each sample was sequenced using Illumina platform. For this experiment, 3 control samples and 3 mutant samples were analysed.本研究旨在探究泪腺上皮中Pea3介导的信号通路下游的转录靶标。为筛选潜在靶标，我们对对照胚胎以及携带上皮特异性Pea3敲除的突变胚胎的胚胎第14.5天（E14.5）泪腺上皮组织实施了激光捕获显微切割（laser capture microdissection, LCM）。组织取材后，提取RNA。采用Clontech SMART-seq v4 超低起始量RNA建库试剂盒完成cDNA合成与扩增，并依托哥伦比亚大学（Columbia University）核心实验平台，使用Nextera XT DNA文库制备试剂盒完成cDNA文库构建，后续进行RNA测序。文库制备完成后，通过Illumina测序平台对所有样本进行测序。本实验共分析3份对照样本与3份突变样本。