A versatile viral toolkit for functional discovery in the nervous system.
A versatile viral toolkit for functional discovery in the nervous system.
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DOI：
10.1016/j.crmeth.2022.100225
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发表时间：
2022-06-20
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精确控制转基因表达的能力对于基础研究和临床应用是必不可少的。腺相关病毒（AAV）是非致病性的，并且可用于在几乎任何组织、细胞类型或物种中驱动稳定表达，但其有限的基因组有效载荷导致可掺入的转基因与控制其表达的调控元件的复杂性之间的权衡。在复杂的实验中解决这些相互竞争的必要性不可避免地会导致妥协。在这里，我们组装了一个优化的病毒工具包（VTK），解决了这些限制，并允许有效的组合靶向细胞类型。此外，它们的模块化设计明确地允许进一步的改进。我们通过将AAV载体的结构改进与创新的分子工具相结合，在紧凑型载体中实现了这一点。我们说明了这种方法的潜力，通过系统的演示其效用为目标的细胞类型和查询其生物学使用广泛的遗传编码的工具。细胞类型特异性靶向是研究电路和行为所必需的因素，如转基因大小限制了可用的AAV有效载荷VTK是一组优化的AAV骨架，克服了这些限制。允许精确标记和操纵神经元的转基因的多样性不断发展，但能够将其表达限制为特定和功能定义的亚型的病毒载体的进化落后了。在这里，我们提出了一种优化的病毒工具包（VTK），旨在通过增加AAV载体的模块化来简化实验设计，以靶向特定类型的神经元。Pouchelon等人开发一种优化的模块化病毒工具包（VTK），其解决了关键的AAV限制并允许神经元细胞类型的有效组合靶向。这种方法允许使用广泛的遗传编码工具来有效地查询神经元生物学。
The ability to precisely control transgene expression is essential for basic research and clinical applications. Adeno-associated viruses (AAVs) are non-pathogenic and can be used to drive stable expression in virtually any tissue, cell type, or species, but their limited genomic payload results in a trade-off between the transgenes that can be incorporated and the complexity of the regulatory elements controlling their expression. Resolving these competing imperatives in complex experiments inevitably results in compromises. Here, we assemble an optimized viral toolkit (VTK) that addresses these limitations and allows for efficient combinatorial targeting of cell types. Moreover, their modular design explicitly enables further refinements. We achieve this in compact vectors by integrating structural improvements of AAV vectors with innovative molecular tools. We illustrate the potential of this approach through a systematic demonstration of their utility for targeting cell types and querying their biology using a wide array of genetically encoded tools. Cell-type-specific targeting is required to study circuits and behaviors Factors such as transgene size limit the AAV payload available VTK is a set of optimized AAV backbones that overcomes these limitations The diversity of transgenes allowing for precise labeling and manipulation of neurons is ever evolving, but the evolution of viral vectors capable of restricting their expression to specific and functionally defined subtypes lags behind. Here, we present an optimized viral toolkit (VTK) that aims at simplifying the experimental design by increasing the modularity of AAV vectors for the targeting of specific types of neurons. Pouchelon et al. develop an optimized and modular viral toolkit (VTK) that addresses key AAV limitations and allows for efficient combinatorial targeting of neuronal cell types. This approach allows efficient querying of neuronal biology using a wide array of genetically encoded tools.