Identification of neuron-type specific promoters in monkey genome and their functional validation in mice.
Identification of neuron-type specific promoters in monkey genome and their functional validation in mice.
复制标题
DOI：
10.1016/j.bbrc.2019.08.101
复制
发表时间：
2019-10
影响因子：
3.1
通讯作者：
Yuma Nagai;N. Nishitani;Masaharu Yasuda;Y. Ueda;Yuto Fukui;Chihiro Andoh;H. Shirakawa;T. Nakagawa;Ken-ichi Inoue;K. Nagayasu;S. Kasparov;Kae Nakamura;S. Kaneko
中科院分区：
文献类型：
--
作者：
Yuma Nagai;N. Nishitani;Masaharu Yasuda;Y. Ueda;Yuto Fukui;Chihiro Andoh;H. Shirakawa;T. Nakagawa;Ken-ichi Inoue;K. Nagayasu;S. Kasparov;Kae Nakamura;S. Kaneko
病毒基因传递是解释脑功能障碍（如神经精神障碍）机制的最通用的技术之一。由于大脑的复杂性，基因工具的表达，如通道视紫红质和钙传感器，通常必须限制在与这些疾病有关的电路中的特定细胞类型。目前只有少数针对神经元亚型的启动子用于病毒基因传递。在这里，我们从猕猴基因组中分离出几个亚型特异性基因的保守启动子区域，并研究了它们在慢病毒载体（LVVs）中使用时在小鼠大脑中的功能。免疫组织化学分析显示，由生长抑素（SST）、胆囊收缩素（CCK）、小白蛋白（PV）、血清素转运蛋白（SERT）、囊泡乙酰胆碱转运蛋白（vAChT）、P物质（SP）和脑啡肽（PENK）启动子序列诱导的转基因表达在很大程度上与靶向神经元群体的特异性标记共定位。此外，通过将这些结果与转录因子与分离序列结合的计算机预测相结合，我们确定了可能具有细胞类型特异性的转录因子。这些发现为扩展当前适合阐明这些神经元表型的启动子工具箱奠定了基础。
Viral gene delivery is one of the most versatile techniques for elucidating the mechanisms underlying brain dysfunction, such as neuropsychiatric disorders. Due to the complexity of the brain, expression of genetic tools, such as channelrhodopsin and calcium sensors, often has to be restricted to a specified cell type within a circuit implicated in these disorders. Only a handful of promoters targeting neuronal subtypes are currently used for viral gene delivery. Here, we isolated conserved promoter regions of several subtype-specific genes from the macaque genome and investigated their functionality in the mouse brain when used within lentiviral vectors (LVVs). Immunohistochemical analysis revealed that transgene expression induced by the promoter sequences for somatostatin (SST), cholecystokinin (CCK), parvalbumin (PV), serotonin transporter (SERT), vesicular acetylcholine transporter (vAChT), substance P (SP) and proenkephalin (PENK) was largely colocalized with specific markers for the targeted neuronal populations. Moreover, by combining these results with in silico predictions of transcription factor binding to the isolated sequences, we identified transcription factors possibly underlying cell-type specificity. These findings lay a foundation for the expansion of the current toolbox of promoters suitable for elucidating these neuronal phenotypes.