Rat liver-targeted naked plasmid DNA transfer by tail vein injection
Rat liver-targeted naked plasmid DNA transfer by tail vein injection
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DOI：
10.1385/mb:26:2:165
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发表时间：
2004-02-01
影响因子：
2.6
通讯作者：
Gejyo, F
中科院分区：
文献类型：
--
作者：
Maruyama, H;Higuchi, N;Gejyo, F
通过“基于流体动力学的转染”，即向尾静脉中快速注射大量裸露的脱氧核糖核酸（DNA）溶液，可以实现小鼠肝细胞中高水平的外源基因表达。与小鼠相比，大鼠对监测血液参数所需的频繁静脉切开术具有更高的耐受性，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裸露的质粒 DNA 传递到正常大鼠体内，正常大鼠的体型比小鼠大 10 倍以上。我们使用配备有外管的带翼针的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 导致在肝脏中独家检测到 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 杯）。当 15 秒内转移 25 mL 注射体积（约 100 mL/kg 体重）时，可实现最大 Epo 基因表达。
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 P-galactosidase in the liver. We also injected a rat erythropoietin (Epo) expression plasmid, pCAGGS-Epo (800 mug). Maximal Epo gene expression was achieved when a 25-mL injection volume (approx 100 mL/kg body wt) was transferred within 15s.