Gene transfer to subdermal tissues via a new gene gun design
Gene transfer to subdermal tissues via a new gene gun design
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DOI：
10.1089/10430340360464732
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发表时间：
2003-01-01
影响因子：
4.2
通讯作者：
Huang, L
中科院分区：
文献类型：
--
作者：
Dileo, J;Miller, TE;Huang, L
尽管粒子介导的基因转移技术（基因枪）已被应用于向外部组织的基因转移，但将该技术应用于其他组织的成功有限。在这里，我们报告了一种新设计的基因枪的发展，使用氦气放电推动DNA涂层的金珠悬浮在液体中。更高的排出压力允许将DNA递送到更深的组织。使用新的基因枪传递荧光素酶表达质粒导致皮肤中的基因表达水平高于传统枪，以及皮下组织，包括皮下肿瘤。即使使用低至125 ng的DNA，皮肤和肌肉中的基因表达在轰击后24小时达到峰值水平，并保持至少1周。LacZ表达质粒的使用表明基因表达分布在整个皮肤中，没有可观察到的病理学。新的基因枪被用于向小鼠递送模型肿瘤排斥抗原（一种修饰的人乳头瘤病毒[HPV] E7基因）。所有接受治疗的动物都产生了针对HPV阳性肿瘤的保护性免疫。这些结果表明，我们的新设计可以用于标准基因枪应用，并将基因枪技术的范围扩展到以前无法获得的组织。
Although particle-mediated gene transfer technology (gene gun) has been applied for gene transfer to external tissues, the application of this technology to other tissues has met with limited success. Here we report the development of a new design of a gene gun that uses helium discharge to propel DNA-coated gold beads that are suspended in liquid. Higher discharge pressures allow for the delivery of DNA to deeper tissues. Using the new gene gun to deliver a luciferase expression plasmid resulted in higher levels of gene expression in the skin than observed with conventional guns, as well as in subdermal tissues, including subcutaneous tumors. Even when using as little as 125 ng of DNA, gene expression in skin and muscle reached its peak level at 24 hr postbombardment and remained for at least 1 week. The use of a LacZ expression plasmid showed that gene expression was distributed throughout the skin with no observable pathology. The new gene gun was used to deliver a model tumor rejection antigen (a modified human papilloma virus [HPV] E7 gene) to mice. All of the treated animals developed protective immunity against HPV-positive tumors. These results demonstrate that our new design can be used in standard gene gun applications and extends the reach of gene gun technology to tissues that were previously unavailable.