Maruyama Hiroki, Higuchi Noboru, Kameda Shigemi, Miyazaki Jun-Ichi, Gejyo Fumitake
Hiroki Maruyama, MD, Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, 1-754 Asahimachi-dori, Niigata 951-8120, Japan.
Mol Biotechnol. 2004 Feb;26(2):165-72. doi: 10.1385/mb:26:2:165.
High levels of foreign gene expression in mouse hepatocytes can be achieved by "hydrodynamics-based transfection," the rapid injection of a large volume of a naked deoxyribonucleic acid (DNA) solution into the tail vein. Rats are more tolerant of the frequent phlebotomies required for monitoring blood parameters than mice and, thus, are more suitable for some biomedical research. Recently, we demonstrated that hydrodynamics-based transfection can also be used to deliver naked plasmid DNA into the normal rat, which is more than 10 times larger than the mouse. We performed the tail vein injection using a syringe with a winged needle equipped with an external tube. Injection of a lac Z expression plasmid, pCAGGS-lac Z by this technique resulted in the exclusive detection of beta-galactosidase in the liver. We also injected a rat erythropoietin (Epo) expression plasmid, pCAGGS-Epo (800 microg). Maximal Epo gene expression was achieved when a 25-mL injection volume (approx 100 mL/kg body wt) was transferred within 15 s.
通过“基于流体动力学的转染”,即将大量无载体脱氧核糖核酸(DNA)溶液快速注入尾静脉,可在小鼠肝细胞中实现高水平的外源基因表达。相比于小鼠,大鼠对监测血液参数所需的频繁采血更耐受,因此更适合某些生物医学研究。最近,我们证明基于流体动力学的转染也可用于将无载体质粒DNA导入正常大鼠体内,大鼠体型比小鼠大十多倍。我们使用配有外置管的翼形针注射器进行尾静脉注射。通过该技术注射lac Z表达质粒pCAGGS-lac Z后,仅在肝脏中检测到β-半乳糖苷酶。我们还注射了大鼠促红细胞生成素(Epo)表达质粒pCAGGS-Epo(800微克)。当在15秒内注入25毫升(约100毫升/千克体重)的注射体积时,可实现最大的Epo基因表达。