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Hydrodynamics-based transfer of PCR-amplified DNA fragments into rat liver

Authors :
Yuji Nishikawa
Shigemi Kameda
Gen Nakamura
Jun-ichi Miyazaki
Hiroki Maruyama
Noboru Higuchi
Noriaki Iino
Fumitake Gejyo
Source :
Biochemical and Biophysical Research Communications. 309:929-936
Publication Year :
2003
Publisher :
Elsevier BV, 2003.

Abstract

A high level of plasmid DNA expression in rat liver can be achieved by the rapid injection of a large volume of a naked DNA solution into the tail vein, called the ‘hydrodynamics-based procedure.’ The preparation of PCR-amplified DNA fragments is easier than that of naked DNA. In this paper we evaluated the effects of expressing the erythropoietin (Epo) gene in the rat liver by injecting fCAGGS-Epo, an Epo-expressing PCR-amplified DNA fragment, via the tail vein. After injection of 5 pmol fCAGGS-Epo (10 μg) or pCAGGS-Epo (18.4 μg), plasmid DNA, the serum Epo levels peaked at week 1, then persisted for at least 12 weeks. Transgene-derived Epo secretion resulted in significant erythropoiesis. These results demonstrated that transfer of PCR-amplified DNA fragments into the rat liver via rapid tail vein injection can be achieved. This method may provide a useful means for studying the physiologic function of a putative gene.

Details

ISSN :
0006291X
Volume :
309
Database :
OpenAIRE
Journal :
Biochemical and Biophysical Research Communications
Accession number :
edsair.doi.dedup.....98b5f51caae8e6430861ce740a052d08