ExBoX – a simple Boolean exclusion strategy to drive expression in neurons

• DOI: 10.1242/jcs.257212
• Publication Date: 2021-09-13T09:17:21Z
• Authors: Teresa Ubina; Tyler Vahedi-Hunter; Will Agnew-Svoboda; Wenny Wong; Akshay Gupta; Vijayalakshmi San...; Martin M. Riccomagno
• Keywords: Allelic exclusion; Cre recombinase

ABSTRACT The advent of modern single-cell biology has revealed the striking molecular diversity cell populations once thought to be more homogeneous. This newly appreciated complexity made intersectional genetic approaches essential understanding and probing cellular heterogeneity at functional level. Here, we build on previous knowledge develop a simple adeno-associated virus (AAV)-based approach define specific subpopulations cells by Boolean exclusion logic (AND NOT). expression (ExBoX) system encodes for gene interest that is turned particular recombinase (Cre or FlpO) off another. ExBoX allows transcription in expressing only activating recombinase, but not both. We show ability tightly regulate fluorescent reporters vitro vivo, further demonstrate adaptability achieving variety virally delivered coding sequences mouse brain. strategy will expand toolkit available cell- time-specific systems.