The resolution and dimensionality with which biologists can characterize cell types have expanded dramatically in recent years, and intersectional consideration of such features (e.g., multiple gene expression and anatomical parameters) is increasingly understood to be essential. At the same time, genetically targeted technology for writing in and reading out activity patterns for cells in living organisms has enabled causal investigation in physiology and behavior; however, cell-type-specific delivery of these tools (including microbial opsins for optogenetics and genetically encoded Ca²⁺ indicators) has thus far fallen short of versatile targeting to cells jointly defined by many individually selected features. Here, we develop a comprehensive intersectional targeting toolbox including 39 novel vectors for joint-feature-targeted delivery of 13 molecular payloads (including opsins, indicators, and fluorophores), systematic approaches for development and optimization of new intersectional tools, hardware for in vivo monitoring of expression dynamics, and the first versatile single-virus tools (Triplesect) that enable targeting of triply defined cell types.

近年来，生物学家表征细胞类型的分辨率和维度已显着扩大，并且人们越来越认识到对这些特征（例如，多基因表达和解剖参数）的交叉考虑是至关重要的。与此同时，用于写入和读出活体细胞活动模式的基因靶向技术使得生理学和行为的因果研究成为可能；然而，这些工具（包括用于光遗传学的微生物视蛋白和基因编码的 Ca ²⁺指示剂）的细胞类型特异性传递迄今为止还不足以对由许多单独选择的特征共同定义的细胞进行多功能靶向。在这里，我们开发了一个全面的交叉靶向工具箱，包括 39 个新颖的载体，用于联合特征靶向递送 13 种分子有效负载（包括视蛋白、指示剂和荧光团）、用于开发和优化新交叉工具的系统方法、用于体内监测的硬件表达动力学，以及第一个多功能单病毒工具（Triplesect），能够靶向三重定义的细胞类型。