The Use of Adenovirus Dodecahedron in the Delivery of an Enzymatic Activity in the Cell.

Biotechnology Research International Pub Date : 2016-01-01 Epub Date: 2016-05-08 DOI:10.1155/2016/5030589
Sumarheni, Benoit Gallet, Pascal Fender
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引用次数: 5

Abstract

Penton-dodecahedron (Pt-Dd) derived from adenovirus type 3 is a symmetric complex of pentameric penton base plus fiber which can be produced in the baculovirus system at a high concentration. The size of Pt-Dd is smaller than the virus, but this virus-like particle (VLP) has the major proteins recognized by specific receptors on the surface of almost all types of cell. In this study, by direct observation with fluorescence microscopy on a fixed and living cell, the intracellular trafficking and localization of Pt-Dd labeled with fluorescence dyes in the cytoplasm of HeLa Tub-GFP showed a rapid internalization characteristic. Subsequently, the linkage of horseradish peroxidase (HRP) with Pt-Dd as the vector demonstrated an efficient system to deliver this enzyme into the cell without interfering its enzymatic activity as shown by biochemical and cellular experiments. These results were supported by additional studies using Bs-Dd or free form of the HRP used as the control. Overall, this study strengthens the potential role of Pt-Dd as an alternative vector for delivering therapeutic agents.

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利用腺病毒十二面体在细胞内传递酶活性。
来源于腺病毒3型的penton -dodecahedron (Pt-Dd)是一种由五聚体penton碱和纤维组成的对称复合体,可以在杆状病毒系统中高浓度产生。Pt-Dd的大小比病毒小,但这种病毒样颗粒(VLP)具有几乎所有类型细胞表面特定受体识别的主要蛋白质。本研究通过在固定细胞和活细胞上的荧光显微镜直接观察,荧光染料标记的Pt-Dd在HeLa Tub-GFP细胞质中的胞内转运和定位表现出快速内化的特点。随后,通过生化和细胞实验证明,辣根过氧化物酶(HRP)与Pt-Dd作为载体的连接是一种有效的系统,可以将该酶传递到细胞中,而不会干扰其酶活性。这些结果得到了使用Bs-Dd或自由形式的HRP作为对照的其他研究的支持。总的来说,这项研究加强了Pt-Dd作为递送治疗剂的替代载体的潜在作用。
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