© 2005 The Japanese Biochemical Society
Regular Paper |
Sustained Transgene Expression in Rat Kidney with Naked Plasmid DNA and PCR-Amplified DNA Fragments
1 Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Niigata 951-8120; 2 Chugai Pharmaceutical Co., Ltd., 2-1-9 Kyobashi, Chuou-ku, Tokyo 104-8301; 3 Division of Gastroenterology, Department of Surgery II, Asahikawa Medical College, 2-1 Midorigaoka-higashi, Asahikawa 078-8510; and 4 Division of Stem Cell Regulation Research, G6, Osaka University Medical School, 2-2 Yamadaoka, Suita 565-0871
* To whom correspondence should be addressed. Phone: +81-25-227-2194, Fax: +81-25-227-0775, E-mail: hirokim{at}med.niigata-u.ac.jp
ABSTRACT
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.