Electroporation in combination with a plasmid vector containing SV40 enhancer elements results in increased and persistent gene expression in mouse muscle
Electroporation in combination with a plasmid vector containing SV40 enhancer elements results in increased and persistent gene expression in mouse muscle
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DOI：
10.1016/s0006-291x(02)02486-5
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发表时间：
2002-11-08
影响因子：
3.1
通讯作者：
Islam, KB
中科院分区：
文献类型：
--
作者：
Blomberg, P;Eskandarpour, M;Islam, KB
通过注射质粒 DNA 将基因转移到肌肉中是可行的，但发生频率较低。然而，通过将质粒 DNA 注射到小鼠肌肉中后使用电穿孔，已经证明基因表达可以增加 150 倍以上。在本次交流中，我们将此技术与含有来自 SV40 增强子的三个 72 bp DNA 元件串联重复的质粒结合使用来研究基因表达。我们的结果表明，电穿孔和携带这些 DNA 元件的质粒载体的组合导致 BALB/c 小鼠肌肉中荧光素酶报告基因的基因表达增加且更持久。基因递送后 14 天，注射并电穿孔携带 SV40 增强子的质粒的肌肉中的基因表达比对照质粒高 16 倍。我们还研究了递送质粒的载体和 DNase 抑制剂金三羧酸 (ATA) 对基因表达的影响。通过将 ATA 与 150 mM 磷酸钠缓冲液结合，与在生理盐水中递送质粒相比，我们能够使基因表达增加 2 倍。这些结果对于开发裸DNA的高效递送技术具有重要意义。 (C) 2002 年爱思唯尔科学（美国）。版权所有。
Gene transfer into muscle upon injection of plasmid DNA is feasible but occurs with low frequency. However, by using electroporation after injection of plasmid DNA into mouse muscle it has been demonstrated that gene expression can be increased more than 150-fold. In this communication, we have used this technique in combination with plasmids containing a tandem repeat of three 72-bp DNA elements from the SV40 enhancer to study gene expression. Our results show that the combination of electroporation and a plasmid vector carrying these DNA elements results in increased and more persistent gene expression of the luciferase reporter gene in BALB/c mouse muscle. At 14 days after gene delivery, the gene expression was 16-fold higher in muscles injected and electroporated with the plasmid carrying the SV40 enhancers than with control plasmid. We have also studied the effects of the vehicle in which the plasmid was delivered, and the DNase inhibitor aurintricarboxylic acid (ATA), on gene expression. By combining ATA with 150 mM sodium phosphate buffer we were able to obtain a 2-fold increase in gene expression compared to delivery of the plasmid in physiological saline. These results are of importance for the development of efficient delivery techniques for naked DNA. (C) 2002 Elsevier Science (USA). All rights reserved.