Recombinant Adenovirus in Neurobiology - Viral Vector Approaches in Neurobiology and Brain Diseases - page 10

Chapter 2

Recombinant Adenovirus in Neurobiology

Qian Li and Tong-Chuan He

Abstract

Recombinant adenoviruses have been extensively applied in basic research and gene therapy. Recently,
there is an increase in neurobiological studies using recombinant adenovirus technology to manipulate
gene expression in neurological systems. Recombinant adenovirus technology provides a useful tool for
investigating the function of a gene of interest, neurocircuitries and the delivery of genes for therapy.
In order to take full advantage of this technique, it is important to understand the strategies for the design
and application of the virus. The goal of this chapter is to provide a practical protocol on the application
of recombinant adenovirus for neurobiological studies. A simplifi ed recombinant adenoviral system,
AdEasy, will be applied to generate recombinant adenoviruses for the expression and knockdown of the
gene of interest. We discuss the construction of vectors for gene expression and siRNAs, the generation of
adenoviral recombinants, and the production of high-titer adenovirus. Furthermore, a procedure for injection
of the adenovirus using a stereotaxic apparatus is discussed.

Key words Adenovirus, siRNA, AdEasy, Brain, Stereotaxic injection

1 Introduction

Recombinant adenovirus has been used for gene therapy for cancer
and other diseases (see reviews [ 1 - 4 ]). On the other hand, the
recombinant adenoviral systems provide useful tools in biomedical
research to manipulate expression of genes of interest both in vivo
and ex vivo. Due to the ease of production and effi cient infection
rate, recombinant adenovirus systems have been widely applied in
the neurobiological studies for determining the function of genes
of interest. For example, we studied the effects of serotonin 1A
(5-HT 1A ) and serotonin 2C (5-HT 2C ) receptors in the amygdala
on anxiety-like behaviors in mice using recombinant adenoviral
systems [ 5 ]. Applications of recombinant adenovirus include over-
expression, knockdown, or inducible knock-in or knockout of
target genes. Injection of recombinant adenovirus containing a
gene of interest into specifi c brain region provides a useful tool
to study the function of the gene of interest in the brain regions.
By including a cell-specifi c promoter in the recombinant adenoviral