High expression of naked plasmid DNA in muscles of young rodents
High expression of naked plasmid DNA in muscles of young rodents
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DOI：
10.1093/hmg/6.9.1435
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发表时间：
1997-09-01
影响因子：
3.5
通讯作者：
Wolff, JA
中科院分区：
文献类型：
--
作者：
Danko, I;Williams, P;Wolff, JA
在小鼠和大鼠中肌内注射裸质粒DMA获得非常高水平的外源基因表达的时间窗在2周龄，使用含有CMV启动子、嵌合内含子和luc（+）荧光素酶基因的构建体在Balb/C小鼠中获得超过1 μ g荧光素酶蛋白/肌肉的最高表达，在2周龄的Balb/C小鼠中肌内注射β-半乳糖苷酶表达载体后，约50%的肌纤维是强烈的蓝色。年龄、小鼠品系和构建体的影响是倍增的，导致>1000倍的荧光素酶和类似于20倍的β-半乳糖苷酶阳性细胞。这些高水平的表达是不稳定的，并且在较大的动物（狗、恒河猴）中没有观察到。这些结果表明，在体内用裸DNA在肌肉中可以获得大量的外源基因表达，并且将使得能够迅速研究啮齿动物肌肉中基因功能和表达的暂时影响。
There is a time window at 2 weeks of age for achieving very high levels of foreign gene expression from the intramuscular injection of naked plasmid DMA in mice and rats, The highest expression, over 1 mu g of luciferase protein/muscle, was obtained in Balb/C mice using constructs containing the CMV promoter, a chimeric intron and the luc(+) luciferase gene, Approximately 50% of the myofibers were intensely blue following the intramuscular injection of a beta-galactosidase expression vector in 2 week old Balb/C mice, The effects of age, mouse strain and construct were multiplicative, resulting in >1000-fold greater luciferase and similar to 20-fold more beta-galactosidase-positive cells. These high levels of expression were unstable and were not observed in larger animals (dog, rhesus monkey), These results indicate that enormous levels of foreign gene expression can be obtained in muscle with naked DNA in vivo and will enable the temporary effects of gene function and expression in rodent muscle to be expeditiously studied.