An Adeno-Associated Virus-Based Toolkit for Preferential Targeting and Manipulating Quiescent Neural Stem Cells in the Adult Hippocampus

Abstract

Summary: Quiescent neural stem cells (qNSCs) with radial morphology are the only proven source of new neurons in the adult mammalian brain. Our understanding of the roles of newly generated neurons depends on the ability to target and manipulate adult qNSCs. Although various strategies have been developed to target and manipulate adult hippocampal qNSCs, they often suffer from prolonged breeding, low recombination efficiency, and non-specific labeling. Therefore, developing a readily manufactured viral vector that allows flexible packaging and robust expression of various transgenes in qNSCs is a pressing need. Here, we report a recombinant adeno-associated virus serotype 4 (rAAV4)-based toolkit that preferentially targets hippocampal qNSCs and allows for lineage tracing, functional analyses, and activity manipulation of adult qNSCs. Importantly, targeting qNSCs in a non-Cre-dependent fashion opens the possibility for studying qNSCs in less genetically tractable animal species and may have translational impact in gene therapy by preferentially targeting qNSCs. : In this article, Song and colleagues report a recombinant adeno-associated virus serotype 4-based toolkit comprising several distinct rAAV4 vectors that preferentially target qNSCs within the hippocampus. They demonstrate that rAAV4-mediated transgene expression is robust in qNSCs and allows for lineage tracing, genetic manipulation using the Cre-Lox system, functional analyses using electrophysiology and calcium imaging, and activity manipulation using chemogenetics. Keywords: adult hippocampal neural stem cells, rAAV4, quiescence, radial, toolkit