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Comparative analysis of the effects of ultrasound-targeted microbubble destruction on recombinant adeno-associated virus- and plasmid-mediated transgene expression in human retinal pigment epithelium cells.

Authors
  • Zheng, Xiao-Zhi
  • Wu, Ying
  • Li, Hong-Li
  • Du, Lian-Fang
  • Wang, Hui-Ping
  • Gu, Qing
Type
Published Article
Journal
Molecular Medicine Reports
Publisher
Spandidos Publications
Publication Date
Jan 01, 2009
Volume
2
Issue
6
Pages
937–942
Identifiers
DOI: 10.3892/mmr_00000195
PMID: 21475924
Source
Medline
License
Unknown

Abstract

Ultrasound-targeted microbubble destruction (UTMD) has been utilized to deliver a drug/gene into cells in both in vitro and in vivo studies. This study was performed to investigate the feasibility of UTMD-enhanced recombinant adeno-associated virus- (rAAV) and plasmid-mediated transfection into the human retinal pigment epithelium (RPE) cell line ARPE-19. Additionally, the transfection efficiency of rAAV and plasmid was compared in order to choose the appropriate gene vector or strategy to be used with UTMD for RPE-based gene modification. rAAV or plasmid encoding an enhanced green fluorescent protein gene was administered to ARPE-19 cells under various UTMD conditions. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl-tetrazolium bromide (MTT) assay. Transfection efficiency was determined by fluorescence microscopy and flow cytometry. UTMD significantly enhanced the transfection efficiency of rAAV and plasmid in ARPE-19 cells without adverse effects on cell viability. The transfection efficiency of rAAV was higher than that of plasmid. rAAV is therefore an appropriate gene vector for use with UTMD in retinal gene modification.

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