BACKGROUND: The growing understanding of the molecular basis of renal diseases makes the development of gene therapy for kidney disorders a potential treatment alternative. Work aimed at determining the feasibility and the efficiency of gene transfer to the kidney using different viral and nonviral transduction systems is a necessary component to understanding the full potential. Lentiviral vectors have been shown to transduce stably different tissues and cell types that are refractory to other gene transfer approaches. To date, the potential of lentiviral vectors to transfer genes in kidney has not been investigated. The scope of this work was to analyze the efficiencies of in vivo transduction of kidney by a lentiviral vector. METHODS: A pseudotyped lentiviral vector carrying the gene for the enhanced green fluorescent protein (EGFP) was delivered into one kidney of experimental mice by retrograde infusion through the ureter. The presence of the virus and the expression of the reporter protein were monitored over time. RESULTS: Both viral DNA and EGFP expression were measurable in the kidney infused with the lentiviral vector but not in the contralateral kidney. Protein expression was detected by immunostaining, as EGFP fluorescence was masked by the high background fluorescence of the kidney. Expression of EGFP persisted for the entire two-month duration of the experiments. CONCLUSIONS: Lentiviral vectors can effectively deliver exogenous genes to the kidney in vivo, resulting in persistent expression of the introduced gene.
BACKGROUND: The growing understanding of the molecular basis of renal diseases makes the development of gene therapy for kidney disorders a potential treatment alternative. Work aimed at determining the feasibility and the efficiency of gene transfer to the kidney using different viral and nonviral transduction systems is a necessary component to understanding the full potential. Lentiviral vectors have been shown to transduce stably different tissues and cell types that are refractory to other gene transfer approaches. To date, the potential of lentiviral vectors to transfer genes in kidney has not been investigated. The scope of this work was to analyze the efficiencies of in vivo transduction of kidney by a lentiviral vector. METHODS: A pseudotyped lentiviral vector carrying the gene for the enhanced green fluorescent protein (EGFP) was delivered into one kidney of experimental mice by retrograde infusion through the ureter. The presence of the virus and the expression of the reporter protein were monitored over time. RESULTS: Both viral DNA and EGFP expression were measurable in the kidney infused with the lentiviral vector but not in the contralateral kidney. Protein expression was detected by immunostaining, as EGFP fluorescence was masked by the high background fluorescence of the kidney. Expression of EGFP persisted for the entire two-month duration of the experiments. CONCLUSIONS: Lentiviral vectors can effectively deliver exogenous genes to the kidney in vivo, resulting in persistent expression of the introduced gene.
Authors: Minjae Kim; Sang Won Park; Mihwa Kim; Sean W C Chen; William T Gerthoffer; Vivette D D'Agati; H Thomas Lee Journal: Am J Physiol Renal Physiol Date: 2010-05-19
Authors: Minjae Kim; Sean W C Chen; Sang Won Park; Mihwa Kim; Vivette D D'Agati; Jay Yang; H Thomas Lee Journal: Kidney Int Date: 2009-02-04 Impact factor: 10.612