Optimized in vivo transfer of small interfering RNA targeting dermal tissue using in vivo surface electroporation.
Optimized in vivo transfer of small interfering RNA targeting dermal tissue using in vivo surface electroporation.
复制标题
DOI：
10.1038/mtna.2012.1
复制
发表时间：
2012-02-14
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
哺乳动物组织的电穿孔 (EP) 是一种已成功应用于临床的技术，用于以 DNA 质粒的形式提供基于基因的疫苗。人们对能够有效地将信使 RNA 和小干扰 RNA (siRNA) 等 RNA 分子递送至哺乳动物组织的平台产生了极大的兴趣。然而，EP 增强的 RNA 体内递送尚未得到广泛表征。本文详细介绍了一种新型低压 EP 方法的电参数优化，该方法可将寡核苷酸（DNA 和 RNA）递送至体内真皮组织。最初，使用这种新型表面真皮 EP 装置优化了编码绿色荧光蛋白 (GFP) 的质粒 DNA 的真皮递送电参数。虽然所有研究的参数都导致可见的转染，但 10V 范围内的电压参数引发了最稳健的信号。然后评估针对 DNA 优化的参数，以将 siRNA 成功电转移至真皮组织。检测到皮肤中强大的标记 siRNA 转染。然后我们评估这些参数是否转化为 siRNA 的成功转移，从而导致体内基因敲低。使用编码 GFP 的报告基因构建体和针对 GFP 信息的标记 siRNA，我们展示了通过 EP 将 siRNA 同时转染到皮肤，并伴随报告基因信号的敲低。 siRNA 递送完成后，没有注射部位炎症或局部组织损伤的证据。因此，微创低压 EP 方法能够以可耐受的方式有效地将 DNA 和 RNA 分子递送至真皮组织。
Electroporation (EP) of mammalian tissue is a technique that has been used successfully in the clinic for the delivery of genetic-based vaccines in the form of DNA plasmids. There is great interest in platforms which efficiently deliver RNA molecules such as messenger RNA and small interfering RNA (siRNA) to mammalian tissue. However, the in vivo delivery of RNA enhanced by EP has not been extensively characterized. This paper details the optimization of electrical parameters for a novel low-voltage EP method to deliver oligonucleotides (both DNA and RNA) to dermal tissue in vivo. Initially, the electrical parameters were optimized for dermal delivery of plasmid DNA encoding green fluorescent protein (GFP) using this novel surface dermal EP device. While all investigated parameters resulted in visible transfection, voltage parameters in the 10 V range elicited the most robust signal. The parameters optimized for DNA, were then assessed for translation of successful electrotransfer of siRNA into dermal tissue. Robust tagged-siRNA transfection in skin was detected. We then assessed whether these parameters translated to successful transfer of siRNA resulting in gene knockdown in vivo. Using a reporter gene construct encoding GFP and tagged siRNA targeting the GFP message, we show simultaneous transfection of the siRNA to the skin via EP and the concomitant knockdown of the reporter gene signal. The siRNA delivery was accomplished with no evidence of injection site inflammation or local tissue damage. The minimally invasive low-voltage EP method is thus capable of efficiently delivering both DNA and RNA molecules to dermal tissue in a tolerable manner.
登录
查看更多内容
影响因子：
5.1
作者：
Broderick, K. E.;Shen, X.;Sardesai, N. Y.
通讯作者：
Sardesai, N. Y.
影响因子：
5.5
作者：
Lin, Feng;Shen, Xuefei;Sardesai, Niranjan Y.
通讯作者：
Sardesai, Niranjan Y.
影响因子：
5.4
作者：
Laddy, Dominick J.;Yan, Jian;Weiner, David B.
通讯作者：
Weiner, David B.
DOI：
10.1016/j.bbrc.2009.06.150
发表时间：
2009-10-23
影响因子：
3.1
作者：
Nakagawa, Shuji;Arai, Yuji;Nakanishi, Tohru
通讯作者：
Nakanishi, Tohru
DOI：
10.1038/nrd2310
发表时间：
2007-06
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
de Fougerolles A;Vornlocher HP;Maraganore J;Lieberman J
通讯作者：
Lieberman J