Proteomic profiling of salivary gland after nonviral gene transfer mediated by conventional plasmids and minicircles.
Proteomic profiling of salivary gland after nonviral gene transfer mediated by conventional plasmids and minicircles.
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DOI：
10.1038/mtm.2014.7
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发表时间：
2014
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影响因子：
--
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中科院分区：
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在这项研究中，我们比较了超声辅助的非病毒基因转移与常规载体和微环载体在小鼠下颌下唾液腺中的基因转移效率和宿主反应。最初，我们观察了等摩尔量的质粒和微环载体的基因转移效率，证实了早期的报告，即在基因转移的物理方法的背景下，微环更有效。然后，我们试图利用全球蛋白质组学图谱来描述唾液腺对外源基因转移的生理反应。有些令人惊讶的是，我们发现，在没有基因转移载体存在的情况下，Sonopation单独对唾液腺蛋白质组几乎没有影响。然而，当使用质粒载体时，我们观察到唾液腺蛋白质组在大剂量腺相关病毒注射后在大小上与先前报告中看到的相比发生了深刻的扰动。最后，我们发现，带有微环的基因转移只会诱导类似于单独使用声波修饰的微小蛋白质组改变。利用质谱仪，我们将蛋白质ID分配给了218个不同于质粒和微环的凝胶点。对这些蛋白质的生物信息学分析表明，它们集中在68条已知的蛋白质相互作用途径上，其中最显著的是与先天免疫、细胞应激和形态发生有关的途径。
In this study, we compared gene transfer efficiency and host response to ultrasound-assisted, nonviral gene transfer with a conventional plasmid and a minicircle vector in the submandibular salivary glands of mice. Initially, we looked at gene transfer efficiency with equimolar amounts of the plasmid and minicircle vectors, corroborating an earlier report showing that minicircle is more efficient in the context of a physical method of gene transfer. We then sought to characterize the physiological response of the salivary gland to exogenous gene transfer using global proteomic profiling. Somewhat surprisingly, we found that sonoporation alone, without a gene transfer vector present, had virtually no effect on the salivary gland proteome. However, when a plasmid vector was used, we observed profound perturbations of the salivary gland proteome that compared in magnitude to that seen in a previous report after high doses of adeno-associated virus. Finally, we found that gene transfer with a minicircle induces only minor proteomic alterations that were similar to sonoporation alone. Using mass spectrometry, we assigned protein IDs to 218 gel spots that differed between plasmid and minicircle. Bioinformatic analysis of these proteins demonstrated convergence on 68 known protein interaction pathways, most notably those associated with innate immunity, cellular stress, and morphogenesis.
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DOI：
10.1073/pnas.1210662109
发表时间：
2012-11-20
影响因子：
11.1
作者：
Baum, Bruce J.;Alevizos, Ilias;Illei, Gabor G.
通讯作者：
Illei, Gabor G.
影响因子：
5.1
作者：
Passineau, M. J.;Zourelias, L.;Benza, R. L.
通讯作者：
Benza, R. L.
影响因子：
5.8
作者：
Zambon, Alexander C.;Gaj, Stan;Salomonis, Nathan
通讯作者：
Salomonis, Nathan
影响因子：
5.1
作者：
Chabot, S.;Orio, J.;Teissie, J.
通讯作者：
Teissie, J.
影响因子：
12.4
作者：
Chen, ZY;He, CY;Kay, MA
通讯作者：
Kay, MA