106 Lentivirus and Adeno‐Associated Virus Mediated Targeted Gene Transfection in Low‐Oxygen Environment

A critical limitation of using retroviral vectors for gene therapy is their inability to infect non‐dividing cells. Although, the adenoviral vectors have the advantage of being able to infect both dividing and non‐dividing cells, they elicit inflammatory response, thus making the interpretation of in vivo experiments harder. Adeno‐associated virus (AAV) and Lentiviral vectors do not have those limitations, however, scant information is available about their transfection efficiency under low‐oxygen tension. To determine if low‐oxygen microenvironment affects viral vector‐mediated gene transfection, we have used two other viral vectors, Adeno‐associated virus (AAV) and Lentiviral constructs in vitro and in vivo to express foreign genes in hypoxic cultured human dermal fibroblasts and ischemic rat wounds. Both cultured normoxic and hypoxic (1% O 2 ) human dermal fibroblasts were identically transfected by the AAV vector. A lenti6‐LacZ construct was injected onto the periphery of rat ischemic and non‐ischemic wound (10 6 pfu/wound) at the time of wounding. Wounds were harvested at post‐operative day 7. Frozen sections of the wounds were fixed in cold acetone and stained with a in situ β‐gal staining kit. Intense expression of β‐gal was observed without any inflammatory response. No significant difference of transfection efficiency was observed between the ischemic and non‐ischemic wounds. Thus our data indicates that both AAV and Lentiviral vectors are suitable to use in gene‐therapy experiments in both ischemic and non‐ischemic cells and tissues in vitro and in vivo.