Cre Activated and Inactivated Recombinant Adeno-Associated Viral Vectors for Neuronal Anatomical Tracing or Activity Manipulation

Curr Protoc Neurosci. 2015 Jul 1;72:1.24.1-1.24.15. doi: 10.1002/0471142301.ns0124s72.

Abstract

Recombinant adeno-associated viruses (rAAVs) transcriptionally activated by Cre recombinase (Cre-On) are powerful tools for determining the anatomy and function of genetically defined neuronal types in transgenic Cre driver mice. Here we describe how rAAVs transcriptionally inactivated by Cre (Cre-Off) can be used in conjunction with Cre-On rAAVs or genomic Cre-reporter alleles to study brain circuits. Intracranial injection of Cre-On/Cre-Off rAAVs into spatially intermingled Cre(+) and Cre(-) neurons allows these populations to be differentially labeled or manipulated within individual animals. This comparison helps define the unique properties of Cre(+) neurons, highlighting the specialized role they play in their constituent brain circuits. This protocol touches on the conceptual and experimental background of Cre-Off rAAV systems, including caveats and methods of validation.

Keywords: AAV; Cre-dependent virus; Cre-lox; rAAV; viral tracer.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Animals
  • Dependovirus / genetics*
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism*
  • Integrases / genetics
  • Integrases / metabolism*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Neurons / metabolism*
  • Transduction, Genetic

Substances

  • Luminescent Proteins
  • Cre recombinase
  • Integrases