Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA, USA.
National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
Recent success in identifying gene-regulatory elements in the context of recombinant adeno-associated virus vectors has enabled cell-type-restricted gene expression. However, within the cerebral cortex these tools are largely limited to broad classes of neurons. To overcome this limitation, we developed a strategy that led to the identification of multiple new enhancers to target functionally distinct neuronal subtypes. By investigating the regulatory landscape of the disease gene Scn1a, we discovered enhancers selective for parvalbumin (PV) and vasoactive intestinal peptide-expressing interneurons. Demonstrating the functional utility of these elements, we show that the PV-specific enhancer allowed for the selective targeting and manipulation of these neurons across vertebrate species, including humans. Finally, we demonstrate that our selection method is generalizable and characterizes additional PV-specific enhancers with exquisite specificity within distinct brain regions. Altogether, these viral tools can be used for cell-type-specific circuit manipulation and hold considerable promise for use in therapeutic interventions.
最近在重组腺相关病毒载体的背景下识别基因调控元件方面的成功，使得细胞类型特异性基因表达成为可能。然而，在大脑皮层中，这些工具在很大程度上仅限于广泛的神经元类别。为了克服这一限制，我们开发了一种策略，从而鉴定了多个新的增强子，以针对功能不同的神经元亚型。通过研究疾病基因 Scn1a 的调控景观，我们发现了针对钙结合蛋白 Parvalbumin (PV) 和血管活性肠肽表达中间神经元的增强子。证明了这些元件的功能实用性，我们表明，PV 特异性增强子允许在包括人类在内的脊椎动物物种中选择性地靶向和操纵这些神经元。最后，我们证明我们的选择方法是可推广的，并在不同的脑区具有高度特异性的额外的 PV 特异性增强子。总之，这些病毒工具可用于细胞类型特异性的回路操作，并在治疗干预中具有很大的应用前景。
