Marked Diversity of Unique Cortical Enhancers Enables Neuron-Specific Tools by Enhancer-Driven Gene Expression.
Marked Diversity of Unique Cortical Enhancers Enables Neuron-Specific Tools by Enhancer-Driven Gene Expression.
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DOI：
10.1016/j.cub.2018.05.015
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发表时间：
2018-07-09
期刊：
影响因子：
--
通讯作者：
Kentros C
中科院分区：
文献类型：
--
作者：
Blankvoort S;Witter MP;Noonan J;Cotney J;Kentros C
了解神经回路功能需要单独解决其组成部分：特定的神经元细胞类型。然而，不仅神经元细胞类型的精确遗传机制仍然模糊，通过转基因技术获得这些神经元细胞类型也仍然难以捉摸。尽管大多数基因在大脑中表达，但绝大多数基因在许多不同类型的神经元中表达，这表明单独的启动子并不足以区分细胞类型。然而，存在数量级更远的控制转录的遗传顺式调节元件（即，增强子），因此我们在小鼠皮质亚区的显微切割样品中筛选增强子活性。这确定了数千种新的推定增强子，其中许多是特定皮质亚区所特有的。含有这种区域特异性增强子的表达构建体的原核注射导致转基因系驱动特异性地在靶向皮质亚区中的不同细胞组中的表达，即使亲本基因的启动子是相对非特异性的。这些数据展示了利用不同神经元细胞类型的遗传机制来开发可能能够靶向任何感兴趣的神经元回路的遗传工具的前景，我们称之为增强子驱动的基因表达（EDGE）。Blankvoort等人鉴定了在显微解剖的皮质亚区中有活性的增强子，发现与在整个器官中发现的相似数量的新颖和独特的推定增强子。他们使用这种增强子来制造对目标大脑区域的神经元具有特异性表达的转基因小鼠，从非特异性基因中获得特异性遗传工具。
Understanding neural circuit function requires individually addressing their component parts: specific neuronal cell types. However, not only do the precise genetic mechanisms specifying neuronal cell types remain obscure, access to these neuronal cell types by transgenic techniques also remains elusive. Whereas most genes are expressed in the brain, the vast majority are expressed in many different kinds of neurons, suggesting that promoters alone are not sufficiently specific to distinguish cell types. However, there are orders of magnitude more distal genetic cis-regulatory elements controlling transcription (i.e., enhancers), so we screened for enhancer activity in microdissected samples of mouse cortical subregions. This identified thousands of novel putative enhancers, many unique to particular cortical subregions. Pronuclear injection of expression constructs containing such region-specific enhancers resulted in transgenic lines driving expression in distinct sets of cells specifically in the targeted cortical subregions, even though the parent gene’s promoter was relatively non-specific. These data showcase the promise of utilizing the genetic mechanisms underlying the specification of diverse neuronal cell types for the development of genetic tools potentially capable of targeting any neuronal circuit of interest, an approach we call enhancer-driven gene expression (EDGE). Blankvoort et al. identify the enhancers active in microdissected cortical subregions, finding similar numbers of novel and unique putative enhancers as found in entire organs. They use such enhancers to make transgenic mice with expression specific to neurons of the targeted brain region, obtaining specific genetic tools from non-specific genes.
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