Broad Institute, Stanley Center for Psychiatric Research, Cambridge, MA 02142, USA.
Harvard Medical School, Blavatnik Institute, Neurobiology, Boston, MA 02115, USA.
Cell Rep Methods. 2022 May 26;2(6):100225. doi: 10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20.
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.
精确控制转基因表达的能力对于基础研究和临床应用至关重要。腺相关病毒 (AAV) 是无致病性的，可以用来驱动几乎任何组织、细胞类型或物种中的稳定表达，但它们有限的基因组载量导致可整合的转基因和控制其表达的调节元件的复杂性之间存在权衡。在复杂的实验中解决这些相互竞争的需求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)，该工具包解决了这些限制，允许对细胞类型进行有效的组合靶向。此外，它们的模块化设计明确允许进一步改进。我们通过将 AAV 载体的结构改进与创新的分子工具集成到紧凑的载体中来实现这一点。我们通过系统地展示它们在使用广泛的遗传编码工具靶向细胞类型和查询其生物学方面的效用来说明这种方法的潜力。
