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Lentiviral gene transfer into the dorsal root ganglion of adult rats
BACKGROUND: Lentivector-mediated gene delivery into the dorsal root ganglion (DRG) is a promising method for exploring pain pathophysiology and for genetic treatment of chronic neuropathic pain. In this study, a series of modified lentivector particles with different cellular promoters, envelope gly...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179738/ https://www.ncbi.nlm.nih.gov/pubmed/21861915 http://dx.doi.org/10.1186/1744-8069-7-63 |
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author | Yu, Hongwei Fischer, Greg Jia, Guangfu Reiser, Jakob Park, Frank Hogan, Quinn H |
author_facet | Yu, Hongwei Fischer, Greg Jia, Guangfu Reiser, Jakob Park, Frank Hogan, Quinn H |
author_sort | Yu, Hongwei |
collection | PubMed |
description | BACKGROUND: Lentivector-mediated gene delivery into the dorsal root ganglion (DRG) is a promising method for exploring pain pathophysiology and for genetic treatment of chronic neuropathic pain. In this study, a series of modified lentivector particles with different cellular promoters, envelope glycoproteins, and viral accessory proteins were generated to evaluate the requirements for efficient transduction into neuronal cells in vitro and adult rat DRG in vivo. RESULTS: In vitro, lentivectors expressing enhanced green fluorescent protein (EGFP) under control of the human elongation factor 1α (EF1α) promoter and pseudotyped with the conventional vesicular stomatitis virus G protein (VSV-G) envelope exhibited the best performance in the transfer of EGFP into an immortalized DRG sensory neuron cell line at low multiplicities of infection (MOIs), and into primary cultured DRG neurons at higher MOIs. In vivo, injection of either first or second-generation EF1α-EGFP lentivectors directly into adult rat DRGs led to transduction rates of 19 ± 9% and 20 ± 8% EGFP-positive DRG neurons, respectively, detected at 4 weeks post injection. Transduced cells included a full range of neuronal phenotypes, including myelinated neurons as well as both non-peptidergic and peptidergic nociceptive unmyelinated neurons. CONCLUSION: VSV-G pseudotyped lentivectors containing the human elongation factor 1α (EF1α)-EGFP expression cassette demonstrated relatively efficient transduction to sensory neurons following direct injection into the DRG. These results clearly show the potential of lentivectors as a viable system for delivering target genes into DRGs to explore basic mechanisms of neuropathic pain, with the potential for future clinical use in treating chronic pain. |
format | Online Article Text |
id | pubmed-3179738 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-31797382011-09-27 Lentiviral gene transfer into the dorsal root ganglion of adult rats Yu, Hongwei Fischer, Greg Jia, Guangfu Reiser, Jakob Park, Frank Hogan, Quinn H Mol Pain Research BACKGROUND: Lentivector-mediated gene delivery into the dorsal root ganglion (DRG) is a promising method for exploring pain pathophysiology and for genetic treatment of chronic neuropathic pain. In this study, a series of modified lentivector particles with different cellular promoters, envelope glycoproteins, and viral accessory proteins were generated to evaluate the requirements for efficient transduction into neuronal cells in vitro and adult rat DRG in vivo. RESULTS: In vitro, lentivectors expressing enhanced green fluorescent protein (EGFP) under control of the human elongation factor 1α (EF1α) promoter and pseudotyped with the conventional vesicular stomatitis virus G protein (VSV-G) envelope exhibited the best performance in the transfer of EGFP into an immortalized DRG sensory neuron cell line at low multiplicities of infection (MOIs), and into primary cultured DRG neurons at higher MOIs. In vivo, injection of either first or second-generation EF1α-EGFP lentivectors directly into adult rat DRGs led to transduction rates of 19 ± 9% and 20 ± 8% EGFP-positive DRG neurons, respectively, detected at 4 weeks post injection. Transduced cells included a full range of neuronal phenotypes, including myelinated neurons as well as both non-peptidergic and peptidergic nociceptive unmyelinated neurons. CONCLUSION: VSV-G pseudotyped lentivectors containing the human elongation factor 1α (EF1α)-EGFP expression cassette demonstrated relatively efficient transduction to sensory neurons following direct injection into the DRG. These results clearly show the potential of lentivectors as a viable system for delivering target genes into DRGs to explore basic mechanisms of neuropathic pain, with the potential for future clinical use in treating chronic pain. BioMed Central 2011-08-23 /pmc/articles/PMC3179738/ /pubmed/21861915 http://dx.doi.org/10.1186/1744-8069-7-63 Text en Copyright ©2011 Yu et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Yu, Hongwei Fischer, Greg Jia, Guangfu Reiser, Jakob Park, Frank Hogan, Quinn H Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title | Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title_full | Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title_fullStr | Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title_full_unstemmed | Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title_short | Lentiviral gene transfer into the dorsal root ganglion of adult rats |
title_sort | lentiviral gene transfer into the dorsal root ganglion of adult rats |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179738/ https://www.ncbi.nlm.nih.gov/pubmed/21861915 http://dx.doi.org/10.1186/1744-8069-7-63 |
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