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Titolo:
Use of polyethylenimine-adenovirus complexes to examine tripler formation in intact cells
Autore:
Hoque, ATMS; Sedelnikova, OA; Luu, AN; Swaim, WD; Panyutin, IG; Baum, BJ;
Indirizzi:
NIDCR, GTTB, NIH, Bethesda, MD 20892 USA NIDCR Bethesda MD USA 20892NIDCR, GTTB, NIH, Bethesda, MD 20892 USA NIH, Dept Nucl Med, Ctr Clin, Bethesda, MD 20892 USA NIH Bethesda MD USA 20892 Dept Nucl Med, Ctr Clin, Bethesda, MD 20892 USA Natl Inst Dent & Craniofacial Res, Cellular Imaging Core Facil, NIH, Bethesda, MD 20892 USA Natl Inst Dent & Craniofacial Res Bethesda MD USA 20892 sda, MD 20892 USA
Titolo Testata:
ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT
fascicolo: 4, volume: 10, anno: 2000,
pagine: 229 - 241
SICI:
1087-2906(200008)10:4<229:UOPCTE>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
HELIX-FORMING OLIGONUCLEOTIDES; MAMMALIAN-CELLS; TRANSCRIPTION ELONGATION; LABELED OLIGONUCLEOTIDES; EPITHELIAL-CELLS; GENE-EXPRESSION; IN-VITRO; DNA; INHIBITION; I-125;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
37
Recensione:
Indirizzi per estratti:
Indirizzo: Baum, BJ NIDCR, GTTB, NIH, Bldg 10,Room 1N113,MSC-1190, Bethesda, MD 20892USA NIDCR Bldg 10,Room 1N113,MSC-1190 Bethesda MD USA 20892 20892 USA
Citazione:
A.T.M.S. Hoque et al., "Use of polyethylenimine-adenovirus complexes to examine tripler formation in intact cells", ANTISENSE N, 10(4), 2000, pp. 229-241

Abstract

Tripler-forming oligonucleotides (TFOs) show potential for sequence-specific DNA binding and inhibition of gene expression. We have applied this antigene strategy using a TFO incorporating an Auger-emitting radionucleotide, I-125, to study the production of double-strand breaks (dsb) in the rat aquaporin 5 (rAQP5) cDNA, I-125-TFO bound to the pCMVrAQP5 plasmid in vitro ina dose-dependent manner and formed stable triplexes up to 65 degrees C andin the presence of 140 mM KCl. Further, I-125-TFO resulted in a predictable dsb when analyzed by Southern hybridization. To deliver TFOs to epithelial cells, we employed I-125-TFO-polyethyleneimine-adenovirus (I-125-TFO-PEI-Ad) complexes. We hypothesized that these complexes would take advantage ofadenoviral characteristics to transfer I-125-TFO to the cell nucleus. Adenovirus-containing complexes brought about greater uptake and nuclear localization of TFOs compared with delivery with I-125-TFO-PEI complexes alone. No significant degradation of I-125-TFO was found after delivery into cells using PEI-Ad complexes and freezing and thawing. We next used PEI-Ad complexes to deliver I-125-TFO and pCMVrAQP5 separately to epithelial cells to determine if triplexes can form de novo within cells, resulting in the specific dsb in the rAQP5 cDNA, After delivery, cell pellets were stored at -80 degrees C for more than 60 days. Thereafter, plasmid DNA was isolated from cells and analyzed for dsb by Southern hybridization, However, none were detected, We conclude that under the experimental conditions employed, effective triplexes, with I-125-TFO and pCMVrAQP5, do not form de novo inside cells.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/09/20 alle ore 08:43:58