TCF20 regulates neurogenesis in mice brain. TCF20 regulates neurogenesis in mice brain

Purpose:To gain a deeper insight into how TCF20 regulates neurogenesis in mice brain, RNA-sequencing (RNA-seq) was performed to analyze the genome-wide changes by TCF20 deletion at E13.5. Methods: Total RNA was extracted from E13.5 telencephalic tissue of TCF20 WT HET KO mice. Then total RNA was quality controlled and quantified using an Agilent 2100 Bioanalyzer. After converting to cDNA and building library, high-throughput sequencing was performed using the Illumina HiSeq 2500 platform in Annoroad Genomics. Results: Approximately approximately one thousand transcripts showed differential expression between the WT and TCF20 KO brain cortex, with a fold change ≥1.5 and p value <0.05. Gene ontology (GO) analysis showed that the down-regulated genes were enriched in the terms related to neurogenesis, neuronal specification, neural differentiation and secretion by cells. Up-regulated genes showed a significant enrichment of terms involved in up regulation of hypersensitivity and up regulation of inflammatory response. These results reflected the importance of TCF20 in cortical neurogenesis. Conclusions: We conclude that RNA-seq based transcriptome characterization would provide a framework for understanding how TCF20 gene contribute to brain cortical development. Overall design: Total RNA was extracted from E13.5 telencephalic tissue of TCF20 WT HET KO mice and subjected to deep sequencing, in triplicate, using Illumina HiSeq 2500.

研究目的：为深入解析TCF20对小鼠大脑神经发生的调控机制，本研究采用RNA测序（RNA-sequencing, RNA-seq）技术，分析了E13.5时点TCF20基因敲除后全基因组范围内的基因表达变化。
实验方法：从TCF20野生型（WT）、杂合型（HET）与敲除型（KO）小鼠的E13.5端脑组织中提取总RNA，随后采用安捷伦2100生物分析仪（Agilent 2100 Bioanalyzer）对总RNA进行质量质控与定量检测。将总RNA反转录为cDNA并构建测序文库后，在诺禾致源（Annoroad Genomics）依托Illumina HiSeq 2500平台完成高通量测序。
实验结果：野生型与TCF20敲除型小鼠大脑皮层组织中，约1000个转录本呈现显著差异表达，其表达倍数变化≥1.5且P值<0.05。基因本体（Gene Ontology, GO）富集分析显示，下调基因显著富集于神经发生、神经元特化、神经分化及细胞分泌相关的功能条目；上调基因则显著富集于超敏反应上调与炎症反应上调相关的功能条目。上述结果证实了TCF20在皮层神经发生过程中的关键作用。
研究结论：本研究表明，基于RNA测序（RNA-seq）的转录组特征分析，可为解析TCF20基因调控大脑皮层发育的分子机制提供研究框架。
整体实验设计：从TCF20野生型、杂合型与敲除型小鼠的E13.5端脑组织中提取总RNA，每组设置3个生物学重复，采用Illumina HiSeq 2500平台进行深度测序。