Transcription profiling by high throughput sequencing of douple (TNFR1-/-/TNFR2-/-) and triple (Zfp36-/-/TNFR1-/-/TNFR2-/-) knockout mouse spleens

Tristetraprolin (TTP) binds to specific AU-rich elements in the 3'UTR of certain transcripts and regulates post-transcriptional gene expression by increasing the rate of mRNA turnover. In this study, we evaluated the effects of TTP deficiency on the overall gene expression of spleen tissue, in order to discover tissue specific targets of TTP under normal physiologic conditions. We utilized "Triple KO" (Zfp36-/-/TNFR1-/-/TNFR2-/-) mice that are deficient in TTP and two TNF receptors and compared the transcriptomic changes to "Double KO" (TNFR1-/-/TNFR2-/-) and WT mice. Spleen mRNA from four WT, four "Double KO", and four "Triple mice" was subjected to RNA-Seq in two phases. All the animals used in this study were males between the ages of 12-14 weeks and were on a mixed (75% C57BL/6NTac, 25% 129/SVEV) background. Examination of splenic gene expression difference between "Triple KO"-WT and "double KO"-WT data setsTristetraprolin (TTP) 可结合特定转录本3'非翻译区（3'UTR）内的富含AU元件（AU-rich elements），并通过加快mRNA周转速率调控转录后基因表达。本研究旨在探明正常生理条件下TTP的组织特异性靶标，故而评估了TTP缺失对脾脏组织整体基因表达的影响。本研究使用了同时缺失TTP与两种TNF受体的三重敲除（Triple KO, Zfp36-/-/TNFR1-/-/TNFR2-/-）小鼠，并将其转录组变化与双敲除（Double KO, TNFR1-/-/TNFR2-/-）小鼠及野生型（WT）小鼠进行对比。我们分两阶段对4只野生型（WT）、4只双敲除及4只三重敲除小鼠的脾脏mRNA进行了RNA测序（RNA-Seq）。本研究所用的全部实验动物均为12~14周龄的雄性小鼠，遗传背景为混合背景（75% C57BL/6NTac, 25% 129/SVEV）。本研究对三重敲除-野生型与双敲除-野生型数据集的脾脏基因表达差异进行了分析。