RT Journal Article
SR Electronic
T1 ExBoX: a simple Boolean exclusion strategy to drive expression in neurons
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.07.28.224691
DO 10.1101/2020.07.28.224691
A1 Tyler Vahedi-Hunter
A1 Teresa Ubina
A1 Akshay Gupta
A1 Wenny Wong
A1 Will Agnew-Svoboda
A1 Vijayalakshmi Santhakumar
A1 Martin M. Riccomagno
YR 2020
UL http://biorxiv.org/content/early/2020/07/30/2020.07.28.224691.abstract
AB The advent of modern single-cell biology has revealed the striking molecular diversity of cell populations once thought to be more homogeneous. This newly appreciated complexity has made intersectional genetic approaches essential to understanding and probing cellular heterogeneity at the functional level. Here we build on previous knowledge to develop a simple AAV-based approach to define specific subpopulations of cells by Boolean exclusion logic (AND NOT). This Expression by Boolean Exclusion (ExBoX) system encodes for a gene of interest which is turned ON by a particular recombinase (Cre or FlpO) and turned OFF by another. ExBoX allows for the specific transcription of a gene of interest in cells expressing only the activating recombinase, but not in cells expressing both. We show the ability of the ExBoX system to tightly regulate expression of fluorescent reporters both in vitro and in vivo, and further demonstrate the powerful adaptability of the system by achieving expression of a variety of virally-delivered coding sequences in the mouse brain. This simple strategy will expand the molecular toolkit available for cell- and time-specific gene expression in a variety of organisms and systems.Competing Interest StatementThe authors have declared no competing interest.HAHemagglutininAAVAdeno-Associated VirusDIODouble-floxed Inverse OrientationDGDouble-floxed Inverse OrientationGOIGene Of Interest