In vivo gene transfer to kidney by lentiviral vector

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In vivo gene transfer to kidney by lentiviral vector G. Luca Gusella, Elena Fedorova, Daniele Marras, Paul E. Klotman, Mary E. Klotman Kidney International Volume 61, Issue 1, Pages S32-S36 (January 2002) DOI: 10.1046/j.1523-1755.2002.0610s1032.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 Fluorescence microscopy of untreated (candf) and HR′PGK/EGFP-infused kidney (a, b,d, ande) at one month. The corticomedullary junction (a–c) and the cortex areas (d–f) are shown. Sections in b and e, from virus-treated kidney, were treated with sodium borohydride as described in the Methods section. The treatment resulted in a general decrease of the autofluorescence without, however, improving the specific signal. Kidney International 2002 61, S32-S36DOI: (10.1046/j.1523-1755.2002.0610s1032.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 Detection of EGFP transgene DNA in lentiviral transduced kidneys. Kidneys were collected one month (lanes 3 and 4) or two months (lanes 5 and 6) after viral infusion. Total DNA was extracted from kidneys infused with HR′PGK/EGFP (lanes 3 and 5) and contralateral kidneys (lanes 4 and 6) and subjected to PCR amplification for EGFP or G3PDH. Lanes 1 and 2 show the reaction products of the negative control, in which the template DNA was omitted, and the positive control, in which a plasmid containing the EGFP gene was used as template, respectively. MW, DNA molecular weight markers. Kidney International 2002 61, S32-S36DOI: (10.1046/j.1523-1755.2002.0610s1032.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 Expression of EGFP in lentiviral transduced kidneys. Sections were prepared from kidneys collected one month (a and b) or two months (c–e) following retrograde lentiviral infusion or PBS infusion. The immunodetection for EGFP showed clear positive staining in the corticomedullary junction of the HR′PGK/EGFP-infused kidneys (b and d). Control contralateral kidneys (a and c) and a PBS-injected kidney (e) were used as control and showed no specific staining. Kidney International 2002 61, S32-S36DOI: (10.1046/j.1523-1755.2002.0610s1032.x) Copyright © 2002 International Society of Nephrology Terms and Conditions