Gene delivery to neurons of the dorsal root ganglia using adeno-associated viral vectors

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 procedures 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 (μl) of a viral vector stock; however, the procedure is complex, invasive, 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, intrathecal delivery does require much larger amounts of viral stock (10–20 μl) and has the disadvantage that viral particles will leak from the cerebrospinal fl uid to the spinal cord and/or peripheral tissues.