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首页> 外文期刊>Gene therapy >A novel doxycycline inducible autoregulatory plasmid which displays 'on'/'off' regulation suited to gene therapy applications.
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A novel doxycycline inducible autoregulatory plasmid which displays 'on'/'off' regulation suited to gene therapy applications.

机译：一种新颖的强力霉素诱导的自调控质粒，其显示适用于基因治疗应用的“ on” /“ off”调节。

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摘要

The development of transcriptionally controlled systems which function in eukaryotic cells are important for achieving regulated gene expression in gene therapy. In this study we combined the components of the tetracycline-inducible system in self-contained retroviral and plasmid vectors. Regulated reporter gene expression from the autoregulatory plasmid pGTRTL in response to doxycycline (Dox) induction surpasses the expression observed from other self-contained retroviral and plasmid vectors. Induction kinetics and expression levels of luciferase and the therapeutic molecule, truncated soluble complement receptor 1 (sCR1) were characterised in a mouse fibroblast and a human neuroblastoma cell line. The regulatory characteristics of the plasmids were shown to be optimal for gene therapy applications, as there was a rapid reduction in expression levels following removal of Dox. Co-transfection of cells with an autoregulatory plasmid and a Dox inducible enhanced green fluorescent protein (EGFP) plasmid demonstrated the feasibility of using this plasmid combination to achieve parallel regulation of two genes of interest in a single cell under the control of Dox. These novel autoregulatory plasmids display the requirements for gene therapy applications in chronic conditions which are remitting/relapsing such as rheumatoid arthritis or multiple sclerosis, where novel protein therapeutics and combination therapies are needed. Gene Therapy (2000) 7, 2061-2070.
机译：在真核细胞中起作用的转录控制系统的发展对于在基因治疗中实现调节的基因表达很重要。在这项研究中，我们将四环素诱导系统的组成部分整合在自成一体的逆转录病毒和质粒载体中。自体调控质粒pGTRTL响应强力霉素（Dox）诱导而调控的报告基因表达超过了其他自成一体的逆转录病毒和质粒载体所观察到的表达。在小鼠成纤维细胞和人神经母细胞瘤细胞系中表征了荧光素酶和治疗分子，截短的可溶性补体受体1（sCR1）的诱导动力学和表达水平。质粒的调控特性显示出对于基因治疗应用而言是最佳的，因为去除Dox后表达水平迅速下降。用自动调节质粒和Dox可诱导的增强型绿色荧光蛋白（EGFP）质粒共转染细胞证明了使用这种质粒组合在Dox的控制下在单个细胞中实现两个目的基因的平行调节的可行性。这些新型的自动调节质粒显示出了在慢性疾病（如风湿性关节炎或多发性硬化症）的慢性病/复发性疾病中基因治疗应用的需求，在这种情况下，需要新型的蛋白质疗法和联合疗法。 Gene Therapy（2000）7，2061-2070。

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