Comparison of AAV Serotypes for Gene Delivery to Dorsal Root Ganglion Neurons

Abstract

For many experiments in the study of the peripheral nervous system, it would be useful to genetically manipulate primary sensory neurons. We have compared vectors based on adeno-associated virus (AAV) serotypes 1, 2, 3, 4, 5, 6, and 8, and lentivirus (LV), all expressing green fluorescent protein (GFP), for efficiency of transduction of sensory neurons, expression level, cellular tropism, and persistence of transgene expression following direct injection into the dorsal root ganglia (DRG), using histological quantification and qPCR. Two weeks after injection, AAV1, AAV5, and AAV6 had transduced the most neurons. The time course of GFP expression from these three vectors was studied from 1 to 12 weeks after injection. AAV5 was the most effective serotype overall, followed by AAV1. Both these serotypes showed increasing neuronal transduction rates at later time points, with some injections of AAV5 yielding over 90% of DRG neurons GFP(+) at 12 weeks. AAV6 performed well initially, but transduction rates declined dramatically between 4 and 12 weeks. AAV1 and AAV5 both transduced large-diameter neurons, IB4(+) neurons, and CGRP(+) neurons. In conclusion, AAV5 is a highly effective gene therapy vector for primary sensory neurons following direct injection into the DRG.

Gene delivery to rat and human Schwann cells and nerve segments: a comparison of AAV 1–9 and lentiviral vectors

“…74 These six amino acids include the K531 residue critical for heparan binding. Similar adverse effects of AAV6 were reported in dorsal root ganglion neurons 75,76 and in hippocampal neurons and astrocytes. 77 A mutant form of AAV6 with an altered heparin-binding site 55 did eliminate the toxicity in primary hippocampal cells.…”

Regenerative medicine for the treatment of spinal cord injury: more than just promises?

“…This “new age generation” of viral vectors could be either applied into stem cells before transplantation is performed ( ex vivo ) or injected directly in situ into the damaged-impaired CNS tissue [21]. In this respect, the adeno-associated viral vectors (AAVs), particularly serotype AAV-2 [85] or AAV-1 and AAV-5 [86], represent one of the most attractive gene delivery systems for targeted gene therapy to the nervous tissue. They are able to efficiently transduce neurons while inducing minimal immune responses in the host brain [85,87,88].…”

A novel analgesic approach to optogenetically and specifically inhibit pain transmission using TRPV1 promoter

“…Therefore we directly injected AAV vectors into the mouse DRG so that the viral infection could be limited locally and protein can be specifically expressed in DRG. Previous study showed that AAV5 has the highest transfection rate following direct injection into the DRG (Mason et al, 2010). Therefore, we used AAV5 as a vector and constructed recombinant virus AAV5-TRPV1-ArchT-eGFP using TRPV1 promoter to express ArchT selectively in DRG neurons following local DRG injection.…”

“…Adeno-associated virus (AAV) is one of the most commonly used gene therapy vectors, which has several advantages including low immunogenicity, absence of toxicity as well as stable and safe gene expression (Mason et al, 2010;Towne et al, 2009). A number of AAV serotypes have been applied to mice.…”