Sustained transgene expression in rat kidney with naked plasmid DNA and PCR-amplified DNA fragments.
Recently, we developed a kidney-targeted gene transfer technique, in which naked DNA was injected into the renal vein while the renal vein and artery were clamped. Kidney-targeted DNA transfer with only the renal vein clamped is an important modification that may permit less invasive catheter-based gene transfer in future clinical applications. The preparation of PCR-amplified DNA fragments is less time-consuming than that of naked plasmid DNA. We examined rat erythropoietin (Epo) plasmid, pCAGGS-Epo, or PCR-amplified DNA fragment, fCAGGS-Epo, transfer into the rat kidney with only the renal vein clamped. The Epo level peaked at week 3 and then was sustained for 24 weeks, which resulted in significant erythropoiesis. This modified technique, allowing long-term expression of both PCR-amplified DNA fragments and naked plasmid DNA, could potentially be used for catheter-based gene transfer in humans, and could help determine the physiological functions of putative genes.
Maruyama
Hiroki
H
Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Niigata 951-8120. hirokim@med.niigata-u.ac.jp
Higuchi
Noboru
N
Kameda
Shigemi
S
Nakamura
Gen
G
Shimotori
Masaaki
M
Iino
Noriaki
N
Higuchi
Masato
M
Neichi
Tomohiro
T
Yokoyama
Sadaaki
S
Kono
Toru
T
Miyazaki
Jun-ichi
J
Gejyo
Fumitake
F
D000818:Animals; D004247:DNA / Q000235:genetics*; D004921:Erythropoietin / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D007668:Kidney / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D008297:Male; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D012082:Renal Veins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*; D019076:Transgenes
D004921:Erythropoietin; D004247:DNA