Comprehensive dual- and triple-feature intersectional single-vector delivery of diverse functional payloads to cells of behaving mammals

Both 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 within 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 Ca(2+) indicators) has thus far fallen short of versatile targeting to cells jointly defined by many individually-selected features. In this Resource, 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.