Proteomic profiling of salivary gland after nonviral gene transfer mediated by conventional plasmids and minicircles

Ramaz Geguchadze, Zhimin Wang, Lee Zourelias, Paola Perez-Riveros, Paul C. Edwards, Laurie Machen, Michael J. Passineau

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

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.

Original languageEnglish (US)
Article number14007
Number of pages1
JournalMolecular Therapy - Methods and Clinical Development
Volume1
DOIs
StatePublished - Jan 8 2014

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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