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首页> 外文期刊>Preparative biochemistry & biotechnology: An international journal for rapid communication >Ultrasound enhances retrovirus-mediated gene transfer
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Ultrasound enhances retrovirus-mediated gene transfer

机译：超声增强逆转录病毒介导的基因转移

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摘要

Viral vector systems are efficient for transfection of foreign genes into many tissues. Especially, retrovirus based vectors integrate the transgene into the genome of the target cells, which can sustain long term expression. However, it has been demonstrated that the transduction efficiency using retrovirus is relatively lower than those of other viruses. Ultrasound was recently reported to increase gene expression using plasmid DNA, with or without, a delivery vehicle. However, there are no reports, which show an ultrasound effect to retrovirus-mediated gene transfer efficiency. Retrovirus-mediated gene transfer systems were used for transfection of 293T cells, bovine aortic endothelial cells (BAECs), rat aortic smooth muscle cells (RASMCs), and rat skeletal muscle myoblasts (L6 cells) with beta-galactosidase (beta-Gal) genes. Transduction efficiency and cell viability assay were performed on 293T cells that were exposed to varying durations (5 to 30 seconds) and power levels (1.0 watts/Cm-2 to 4.0 watts/cm(2)) of ultrasound after being transduced by a retrovirus. Effects of ultrasound to the retrovirus itself was evaluated by transduction efficiency of 293T cells. After exposure to varying power levels of ultrasound to a retrovirus for 5 seconds, 293T cells were transduced by a retrovirus, and transduction efficiency was evaluated. Below 1.0 watts/cm(2) and 5 seconds exposure, ultrasound showed increased transduction efficiency and no cytotoxicity to 293T cells transduced by a retrovirus. Also, ultrasound showed no toxicity to the virus itself at the same condition. Exposure of 5 seconds at the power of 1.0 watts/cm(2) of an ultrasound resulted in significant increases in retrovirus-mediated gene expression in all four cell types tested in this experiment. Transduction efficiencies by ultrasound were enhanced 6.6-fold, 4.8-fold, 2.3-fold, and 3.2-fold in 293T cells, BAECs, RASMCs, and L6 cells, respectively. Furthermore, G-Gal activities were also increased by the retrovirus with ultrasound exposure in these cells. Adjunctive ultrasound exposure was associated with enhanced retrovirus-mediated transgene expression in vitro. Ultrasound associated local gene therapy has potential for not only plasmid-DNA-, but also retrovirus-mediated gene transfer.
机译：病毒载体系统对于将外源基因转染到许多组织中是有效的。特别地，基于逆转录病毒的载体将转基因整合到靶细胞的基因组中，其可以维持长期表达。但是，已经证明，使用逆转录病毒的转导效率相对低于其他病毒。最近报道了在有或没有递送载体的情况下，超声使用质粒DNA增加基因表达。但是，没有报道显示超声对逆转录病毒介导的基因转移效率有影响。逆转录病毒介导的基因转移系统用于转染293T细胞，牛主动脉内皮细胞（BAEC），大鼠主动脉平滑肌细胞（RASMC）和大鼠骨骼肌成肌细胞（L6细胞）和β-半乳糖苷酶（β-Gal）基因。在逆转录病毒转导后暴露于不同持续时间（5至30秒）和功率水平（1.0瓦特/ Cm-2至4.0瓦特/ cm（2））的293T细胞上进行了转导效率和细胞活力测定。超声对逆转录病毒本身的影响通过293T细胞的转导效率进行评估。在将不同功率的超声波暴露于逆转录病毒5秒钟后，逆转录病毒转导了293T细胞，并评估了转导效率。低于1.0瓦特/厘米（2）和暴露5秒，超声显示转导效率提高，并且对逆转录病毒转导的293T细胞无细胞毒性。同样，超声波在相同条件下对病毒本身没有毒性。在1.0瓦特/厘米（2）的超声功率下暴露5秒导致在此实验中测试的所有四种细胞类型中逆转录病毒介导的基因表达显着增加。在293T细胞，BAEC，RASMC和L6细胞中，超声的转导效率分别提高了6.6倍，4.8倍，2.3倍和3.2倍。此外，逆转录病毒通过在这些细胞中进行超声波暴露，还可以增加G-Gal活性。辅助超声暴露与体外逆转录病毒介导的转基因表达增强有关。超声相关的局部基因治疗不仅具有质粒DNA的潜力，而且具有逆转录病毒介导的基因转移的潜力。

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