Whole-genome gene expression profiling of Pik3cg-depleted mice

We performed whole-genome gene expression profiling in Pik3cg-/- mice and subsequent gene ontology clustering of differentially expressed genes compared to wild type mice, in order to investigate the role of Pik3cg in platelet membrane biogenesis and blood coagulation. Pik3cg-deficient mice were obtained from sources previously described (T. Sasaki et al, 2000, Science 287, 1040-1046), backcrossed onto the C57BL/6J Jax genetic background for eight generations (B6J;129-Pik3cg-tm1Pngr) and then maintained as a closed colony by intercrossing from within the colony (C57BL/6J Jax contribution: 99.6%). Whole blood from three Pik3cg-/- and three C57BL/6J Jax wild type mice (all females, age: 13-16 weeks, Mouse Breeders Diet (Lab Diets 5021-3)) was collected from terminally anaesthetized mice via the retro-orbital sinus. For each RNA extraction, in vitro transcription (IVT) reactions were performed in duplicates.本研究对Pik3cg基因敲除（Pik3cg-/-）小鼠开展全基因组基因表达谱分析，并针对与野生型小鼠相比获得的差异表达基因进行基因本体（Gene Ontology, GO）聚类，以探究Pik3cg在血小板膜生物发生与凝血过程中的作用。Pik3cg缺陷小鼠获取自此前已报道的实验资源（T. Sasaki等，2000，《科学》(Science) 287卷，1040-1046页），将其回交至C57BL/6J Jax遗传背景达8代（品系标识：B6J;129-Pik3cg-tm1Pngr），随后通过种群内近交维持为封闭群（C57BL/6J Jax遗传贡献占比：99.6%）。采集3只Pik3cg-/-小鼠与3只C57BL/6J Jax野生型小鼠的全血：所有受试小鼠均为雌性，周龄13~16周，饲喂小鼠繁育专用饲料（Lab Diets 5021-3），通过眶静脉窦采集终末麻醉小鼠的血液。针对每一次RNA提取操作，均设置2个平行的体外转录（in vitro transcription, IVT）反应。