Hydrodynamics-based transfer of PCR-amplified DNA fragments into rat liver
Hydrodynamics-based transfer of PCR-amplified DNA fragments into rat liver
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DOI：
10.1016/j.bbrc.2003.08.087
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发表时间：
2003-10-03
影响因子：
3.1
通讯作者：
Gejyo, F
中科院分区：
文献类型：
--
作者：
Kameda, S;Maruyama, H;Gejyo, F
将大量裸DNA溶液快速注射到大鼠尾静脉中，可以实现质粒DNA在大鼠肝脏中的高水平表达，称为“基于流体动力学的程序”。PCR扩增的DNA片段的制备比裸DNA的制备容易。在本文中，我们评估的影响，表达促红细胞生成素（Epo）基因在大鼠肝脏中通过注射fCAGGS-Epo，表达Epo的PCR扩增的DNA片段，通过尾静脉。注射5 pmol fCAGGS-Epo（10 μ g）或pCAGGS-Epo（18.4 μ g）质粒DNA后，血清Epo水平在第1周达到峰值，然后持续至少12周。转基因衍生的Epo分泌导致显著的红细胞生成。这些结果表明，可以实现通过快速尾静脉注射将PCR扩增的DNA片段转移到大鼠肝脏中。该方法为研究推测基因的生理功能提供了一种有用的手段。(C)2003年爱思唯尔公司All rights reserved.
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 mug) or pCAGGS-Epo (18.4 mug), 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. (C) 2003 Elsevier Inc. All rights reserved.