| Dokumentenart: | Artikel | ||||
|---|---|---|---|---|---|
| Titel eines Journals oder einer Zeitschrift: | Investigative Opthalmology & Visual Science | ||||
| Verlag: | ASSOC RESEARCH VISION OPHTHALMOLOGY INC | ||||
| Ort der Veröffentlichung: | ROCKVILLE | ||||
| Band: | 59 | ||||
| Nummer des Zeitschriftenheftes oder des Kapitels: | 1 | ||||
| Seitenbereich: | S. 298 | ||||
| Datum: | 2018 | ||||
| Institutionen: | Biologie und Vorklinische Medizin > Institut für Anatomie Biologie und Vorklinische Medizin > Institut für Anatomie > Lehrstuhl für Humananatomie und Embryologie Biologie und Vorklinische Medizin > Institut für Anatomie > Lehrstuhl für Humananatomie und Embryologie > Prof. Dr. Ernst Tamm | ||||
| Identifikationsnummer: |
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| Stichwörter / Keywords: | PROTEIN-KINASE INHIBITOR; DOMINANT-NEGATIVE RHOA; OUTFLOW FACILITY; GENE-TRANSFER; TRANSGENE EXPRESSION; INTRAOCULAR-PRESSURE; UVEOSCLERAL OUTFLOW; CULTURED HUMAN; OPHTHALMIC SOLUTION; ANTERIOR SEGMENTS; trabecular meshwork; FIV vector; MG132; gene therapy proteasomes | ||||
| Dewey-Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie | ||||
| Status: | Veröffentlicht | ||||
| Begutachtet: | Ja, diese Version wurde begutachtet | ||||
| An der Universität Regensburg entstanden: | Ja | ||||
| Dokumenten-ID: | 47681 |
Web of ScienceZusammenfassung
PURPOSE: To determine if proteasome inhibition using MG132 increased the efficiency of FIV vector-mediated transduction in human trabecular meshwork (TM)-1 cells and monkey organ-cultured anterior segments (MOCAS). METHODS: TM-1 cells were pretreated for 1 hour with 0.5% dimethyl sulfoxide (DMSO; vehicle control) or 5 to 50 mu M MG132 and transduced with FIV.GFP (green fluorescent protein)- or ...
Zusammenfassung
PURPOSE: To determine if proteasome inhibition using MG132 increased the efficiency of FIV vector-mediated transduction in human trabecular meshwork (TM)-1 cells and monkey organ-cultured anterior segments (MOCAS). METHODS: TM-1 cells were pretreated for 1 hour with 0.5% dimethyl sulfoxide (DMSO; vehicle control) or 5 to 50 mu M MG132 and transduced with FIV.GFP (green fluorescent protein)- or FIV.mCherry-expressing vector at a multiplicity of transduction (MOT) of 20. At 24 hours, cells were fixed and stained with antibodies for GFP, and positive cells were counted, manually or by fluorescence-activated cell sorting (FACS). Cells transduced with FIV.GFP particles alone were used as controls. The effect of 20 mu M MG132 treatment on high- and low-dose (2 x 10(7) and 0.8 x 10(7) transducing units [TU], respectively) FIV.GFP transduction with or without MG132 was also evaluated in MOCAS using fluorescence microscopy. Vector genome equivalents in cells and tissues were quantified by quantitative (q)PCR on DNA. RESULTS: In the MG132 treatment groups, there was a significant dose-dependent increase in the percentage of transduced cells at all concentrations tested. Vector genome equivalents were also increased in TM-1 cells treated with MG132. Increased FIV.GFP expression in the TM was also observed in MOCAS treated with 20 mu M MG132 and the high dose of vector. Vector genome equivalents were also significantly increased in the MOCAS tissues. Increased transduction was not seen with the low dose of virus. CONCLUSIONS: Proteasome inhibition increased the transduction efficiency of FIV particles in TM-1 cells and MOCAS and may be a useful adjunct for delivery of therapeutic genes to the TM by lentiviral vectors.
Metadaten zuletzt geändert: 28 Jul 2021 17:28
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