Gene Delivery to Neurons of the Dorsal Root Ganglia Using Adeno-Associated Viral Vectors - Gene Delivery and Therapy for Neurological Disorders - page 175

Chapter 7

Gene Delivery to Neurons of the Dorsal Root Ganglia
Using Adeno-Associated Viral Vectors

Nitish D. Fagoe , Ruben Eggers , Joost Verhaagen ,
and Matthew R. J. Mason

Abstract

Viral vector-mediated gene transfer, especially using adeno-associated viral (AAV) vectors, is a powerful
strategy to manipulate gene expression in vivo in injured neurons. In this chapter we provide two methods
to effi ciently transduce dorsal root ganglion (DRG) neurons in vivo using AAV. We describe detailed pro-
cedures to perform direct injections into specifi c DRG and delivery via the intrathecal space to transduce
the lumbar DRG. Finally, we discuss the specifi c advantages and disadvantages of these two methods of
delivery. The main advantages of direct injection are that high transduction rates can be achieved in specifi c
ganglia (L4/L5) with low amounts ( ʼ l) of a viral vector stock; however, the procedure is complex, inva-
sive, and time-consuming. Intrathecal injection has the advantage of being a fast and simple method to
transduce multiple DRG bilaterally, and involves no surgical manipulation of the DRG. However, intrathe-
cal delivery does require much larger amounts of viral stock (10-20 ʼ l) and has the disadvantage that viral
particles will leak from the cerebrospinal fl uid to the spinal cord and/or peripheral tissues.

Key words Dorsal root ganglia, Peripheral nerve injury, Gene delivery, Adeno-associated viral vector

1 Introduction

The use of gene therapy techniques for the study of the nervous
system is nowadays becoming a standard part of many molecular
neurobiology laboratories' toolkit. The current generation of viral
vectors, primarily AAV and LV, are reliable, relatively easy to make
and are well characterized in terms of transduction profi les, and are
known to have low cytotoxicity and immunogenicity. Furthermore,
these vectors are making the transition from bench to bedside as
they are currently being tested in numerous clinical trials in many
types of disease including neurological diseases [ 1 , 2 ]. This makes
them all the more attractive vehicles with which to develop thera-
pies in in vivo models as well as for their application in basic sci-
ence. Viral vectors, especially AAV, are of particular interest for
gene delivery to the nervous system as they effi ciently transduce