In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy

Durable and stratum-specific gene expression in epidermis

“…20 Repeated examination of cornified cells by tape stripping showed persistent green fluorescing cells for both LZRS-EGFP and INV/LTR-EGFP. At 20 weeks after transduction, when mouse skin has turned over at least 20 times, 21 biopsies were obtained from the transduced areas, fixed and cryosectioned.…”

“…In mouse interfollicular epidermis, a stem cell and its descendant transit amplifying cell and the subsequent terminally differentiated cells are organized in a spatially distinct unit, called epidermal proliferative units. 20,21 In the majority of transduced epidermal proliferating units, transcription of the modified LTR appeared to be activated in parallel to the endogenous involucrin promoter. However, in some instances the activity of the hybrid promoter was not tightly regulated.…”

“…20 On day 3 after abrasion, 20-l aliquots of concentrated viruses (containing 2 × 10 7 transducing units) were injected into the hyperproliferative skin. Initial EGFP expression in the skin was assessed by tape stripping the cornified cells and examining the tapes under fluorescent microscopy.…”

Suicide gene therapy for premalignant disease: a new strategy for the treatment of intraepithelial neoplasia

“…This could be accomplished by direct intralesional injection of retroviral vectors, which has been previously been shown to result in sustained gene expression in a large number of transduced cells in vivo. 27 In addition, since transformed cells may not have a high degree of gap junctional intercellular communication, 28 HSV-TK may be of limited use in the treatment of these incipient lesions. CD-mediated bystander effect has previously been shown to be more effective than that of HSV-TK in both cell culture 29 and implanted tumors.…”

Tissue-specific expression and long-term deposition of human collagen VII in the skin of transgenic mice: implications for gene therapy

“…Expression of the reporter transgene was noted in immune tolerant mice up to 16 weeks but persisted only to 3 weeks in immune competent mice. 17 As a first step towards developing a gene therapy approach to DEB, we have used transgenesis in mice to establish a model to test the feasibility of achieving tissue-specific and stable expression of human collagen VII by gene transfer. A transgenic mouse approach is especially important because it is possible to determine if the regulatory sequences of the gene vector are sufficient to drive appropriate tissue-specific expression.…”