title: Manipulating gene expression in projection-specific neuronal populations using combinatorial viral approaches.
abstract: The mammalian brain contains tremendous structural and genetic complexity that is vital for its function. The elucidation of gene expression profiles in the brain, coupled with the development of large-scale connectivity maps and emerging viral vector-based approaches for target-selective gene manipulation, now allow for detailed dissection of gene-circuit interfaces. This protocol details how to perform combinatorial viral injections to manipulate gene expression in subsets of neurons interconnecting two brain regions. This method utilizes stereotaxic injection of a retrograde transducing CAV2-Cre virus into one brain region, combined with injection of a locally transducing Cre-dependent AAV virus into another brain region. This technique is widely applicable to the genetic dissection of neural circuitry, as it enables selective expression of candidate genes, dominant-negatives, fluorescent reporters, or genetic tools within heterogeneous populations of neurons based upon their projection targets.
authors.affiliation: Department of Pharmacology and Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, WA.
authors.forename: Bryan B
authors.initials: BB
authors.lastname: Gore
authors.affiliation: Department of Pharmacology and Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, WA.
authors.forename: Marta E
authors.initials: ME
authors.lastname: Soden
authors.affiliation: Department of Pharmacology and Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, WA.
authors.forename: Larry S
authors.initials: LS
authors.lastname: Zweifel
mesh_terms: D006516:Adenoviruses, Canine; D000818:Animals; D001921:Brain / Q000502:physiology*; D000229:Dependovirus; D015870:Gene Expression / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors; D051379:Mice; D023421:Models, Animal; D009474:Neurons / Q000502:physiology; D019076:Transgenes
keywords: AAV; CAV; neural circuit; stereotaxic surgery; viral vector