Downstream targeted genes of Satb2 regulate synaptic formation and axonogenesis in developing cerebral cortex [RNA-seq]

Special AT-rich sequencebinding protein 2 (SATB2) is essential for the development of cerebral cortex and key molecular node for the establishment of proper neural circuitry and function. Mutations in SATB2 gene lead to SATB2-associated syndrome (SAS), which is characterized by abnormal development of skeleton and central nervus system. We generated Satb2 knockout mouse model through CRISPR-Cas9 technology and performed RNA-seq and ChIP-seq of embryonic cerebral cortex. We conducted RT-qPCR, western blot, immunofluorescence staining, luciferase reporter assay and behavioral analysis for experimental verification. Comparative gene expression profiling analysis of RNA-seq data for E17.5 wildtype and Satb2 knock-out mouse cerebral cortex特异性AT富集序列结合蛋白2（Special AT-rich sequence binding protein 2，SATB2）对大脑皮层发育不可或缺，同时是构建正常神经环路、维持神经功能的关键分子节点。SATB2基因发生突变可引发SATB2相关综合征（SAS），该病症以骨骼与中枢神经系统发育异常为主要特征。本研究借助CRISPR-Cas9技术构建了Satb2基因敲除小鼠模型，并对胚胎期大脑皮层开展了RNA测序（RNA-seq）与染色质免疫共沉淀测序（ChIP-seq）实验；同时通过实时定量聚合酶链式反应（RT-qPCR）、蛋白质免疫印迹（western blot）、免疫荧光染色、荧光素酶报告基因检测（luciferase reporter assay）以及行为学分析完成实验验证，并针对E17.5野生型与Satb2基因敲除小鼠大脑皮层的RNA-seq数据开展比较基因表达谱分析。