Gene electrotransfer enhanced by nanosecond pulsed electric fields.
Gene electrotransfer enhanced by nanosecond pulsed electric fields.
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DOI：
10.1038/mtm.2014.43
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发表时间：
2014
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影响因子：
--
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纳秒脉冲电场（nsPEFs）对基因电转移的影响在以往的研究中尚未得到明确的证明。本研究旨在评估nsPEFs对编码荧光素酶或绿色荧光蛋白的质粒在HACAT角质形成细胞中的传递及其随后表达的影响。在有或没有nspf的情况下，使用毫秒电脉冲（msep）进行输送。与文献报道相反，我们发现，首先将nsPEFs应用于细胞，然后再应用一个msEP，基因表达显著增加了40倍，而不是相反的顺序。我们证明了nsPEFs对基因转染的影响是有时间限制的。在nsPEFs后立即施用1 msEP可以增强基因表达，并在处理后5分钟达到最大值，在15分钟时略有下降，在1小时时有残余效应。这表明nsPEFs在不改变msep导致的基因表达动力学趋势的情况下发挥了放大器的作用。nspf对细胞活力的影响也取决于特定的脉冲参数。我们还确定了nsPEF对基因电转移的影响涉及钙独立和依赖机制。
The impact of nanosecond pulsed electric fields (nsPEFs) on gene electrotransfer has not been clearly demonstrated in previous studies. This study was conducted to evaluate the influence of nsPEFs on the delivery of plasmids encoding luciferase or green fluorescent protein and subsequent expression in HACAT keratinocyte cells. Delivery was performed using millisecond electric pulses (msEPs) with or without nsPEFs. In contrast to reports in the literature, we discovered that gene expression was significantly increased up to 40-fold by applying nsPEFs to cells first followed by one msEP but not in the opposite order. We demonstrated that the effect of nsPEFs on gene transfection was time restricted. The enhancement of gene expression occurred by applying one msEP immediately after nsPEFs and reached the maximum at posttreatment 5 minutes, slightly decreased at 15 minutes and had a residual effect at 1 hour. It appears that nsPEFs play a role as an amplifier without changing the trend of gene expression kinetics due to msEPs. The effect of nsPEFs on cell viability is also dependent on the specific pulse parameters. We also determined that both calcium independent and dependent mechanisms are involved in nsPEF effects on gene electrotransfer.
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影响因子：
1.5
作者：
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