Key Physiological Phenomena Governing Transgene Expression Based on Tissue Pressure-Mediated Transfection in Mice
Key Physiological Phenomena Governing Transgene Expression Based on Tissue Pressure-Mediated Transfection in Mice
复制标题
DOI：
10.1248/bpb.33.1627
复制
发表时间：
2010-09-01
影响因子：
2
通讯作者：
Hashida, Mitsuru
中科院分区：
文献类型：
--
作者：
Mukai, Hidefumi;Kawakami, Shigeru;Hashida, Mitsuru
体内基因转染是后基因组时代最重要的技术之一。其中，裸质粒DNA转染由于其制备和操作方便以及转染剂无毒性等优点而备受关注。我们已经研究了组织压力介导的转染进行光和控制压力的靶组织后，正常的质粒DNA静脉注射。到目前为止，我们已经证明，质粒DNA和小干扰RNA（siRNA）非常有效地转染到小鼠肾脏，肝脏和脾脏，而不会造成显着的组织损伤。在本研究中，为了了解影响转基因表达的关键生理现象，我们进行了一组涉及组织压力介导转染的实验，包括质粒DNA的生物分布和细胞转运以及转录因子的激活，并获得了以下结果：i）向靶组织及其细胞的质粒DNA转移增加，尽管与施用的总质粒DNA相比，转移的分数很小，ii）诱导质粒DNA的细胞易位的瞬时增加，和iii）激活转录因子。考虑到所有这些结果，似乎组织压力介导的转染增强了质粒DNA向靶组织及其细胞的转移以及转录过程的激活。这一信息将允许更好地理解在体内转基因表达的基础上裸质粒DNA转染，涉及组织压力介导的转染。
It is generally recognized that in viva gene transfection is one of the most important techniques used in the post-genome era. Above all, naked plasmid DNA transfection has attracted much attention because of its advantages including convenience of preparation and handling and lack of toxicity associated with the transfection agents. We have investigated tissue pressure-mediated transfection performed by light and controlled pressure of the target tissue after normal intravenous injection of plasmid DNA. So far, we have demonstrated that plasmid DNA and small-interfering RNA (siRNA) are very efficiently transfected into murine kidney, liver and spleen without causing marked tissue damage. In this study, in order to understand the key physiological phenomena affecting transgene expression, we performed a set of experiments involving tissue pressure-mediated transfection, including the biodistribution and cellular transport of plasmid DNA and activation of transcriptional factors and obtained the following results: i) plasmid DNA transfer to the target tissue and its cells increased although the transferred fraction was small compared to the total administered plasmid DNA, ii) a transient increase in cellular translocation of plasmid DNA was induced, and iii) transcriptional factors were activated. Taking all these results into consideration, it would appear that tissue pressure-mediated transfection enhances plasmid DNA transfer to the target tissue and its cells and also activation of the transcriptional process. This information will allow a better understanding of in viva transgene expression based on naked plasmid DNA transfection involving tissue pressure-mediated transfection.