Reactivation of Silenced Transgene Expression in Mouse Liver by Rapid, Large-Volume Injection of Isotonic Solution
Reactivation of Silenced Transgene Expression in Mouse Liver by Rapid, Large-Volume Injection of Isotonic Solution
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DOI：
10.1089/hum.2008.020
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发表时间：
2008-10-01
影响因子：
4.2
通讯作者：
Takakura, Yoshinobu
中科院分区：
文献类型：
--
作者：
Nishikawa, Makiya;Nakayama, Ayumi;Takakura, Yoshinobu
快速，大容量注射，或所谓的流体动力学注射，裸质粒DNA在小鼠肝脏中具有高的转基因表达，并且该方法已应用于肝脏定向基因转移，仅进行了轻微修改。为了证明注射诱导的生物学变化与水动力注射诱导的小鼠肝脏高水平转基因表达有关，我们将等渗溶液注射到接受水动力注射质粒DNA的小鼠体内。通过这种注射，无论cDNA、启动子和溶液类型如何，肝脏中的转基因表达都增加了。即使在基因转移3个月后，这种再激活也是可重复和可检测的。再激活所需的参数与质粒DNA流体动力学注射所需的参数相似。基于质粒dna -聚乙烯亚胺复合物的转基因在小鼠肝脏中的表达也被同样的处理重新激活。DNA芯片和定量RT-PCR分析显示，立即早期反应基因c-fos和c-jun的表达分别增加了70倍和100倍。激活蛋白(AP)-1或核因子(NF)- κ b依赖的转基因表达通过注射等渗溶液增加。这些发现首次表明，快速、大体积注射等渗溶液可以激活肝脏中的转录因子AP-1和NF-kappa B，从而增加通过流体动力注射或其他方法传递的基因的转录。
Rapid, large-volume injection, or so-called hydrodynamic injection, of naked plasmid DNA gives high transgene expression in mouse liver, and this method has been applied to liver-directed gene transfer in humans with slight modifications. To prove that injection-induced biological changes are involved in hydrodynamic injection-induced, high-level transgene expression in mouse liver, isotonic solutions were injected into mice that had received a hydrodynamic injection of plasmid DNA. Transgene expression in the liver was increased by such injections irrespective of the cDNA, promoter, and type of solution. This reactivation was repeatable and detectable even 3 months after gene transfer. Parameters required for reactivation were similar to those required for the hydrodynamic injection of plasmid DNA. Plasmid DNA-polyethyleneimine complex-based transgene expression in mouse liver was also reactivated by the same treatment. DNA microarray and quantitative RT-PCR analyses revealed that the expression of immediate-early response genes c-fos and c-jun was increased 70-and 100-fold, respectively. Activator protein (AP)-1-or nuclear factor ( NF)-kappa B-dependent transgene expression was increased by an injection of isotonic solutions. These findings indicate for the first time that a rapid, large-volume injection of isotonic solution activates the transcription factors AP-1 and NF-kappa B in the liver, which in turn increases the transcription of genes delivered by hydrodynamic injection or other methods.