Stable expression and cell-specific chromatin structure of human alpha1-antitrypsin cosmid transgenes in rat hepatoma cells.

Détails

ID Serval
serval:BIB_93D521755503
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Stable expression and cell-specific chromatin structure of human alpha1-antitrypsin cosmid transgenes in rat hepatoma cells.
Périodique
Nucleic Acids Research
Auteur(s)
Rollini P., Xu L., Fournier R.E.
ISSN
1362-4962 (Electronic)
ISSN-L
0305-1048
Statut éditorial
Publié
Date de publication
2000
Volume
28
Numéro
18
Pages
3605-3614
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Résumé
The human gene encoding alpha1-antitrypsin (alpha1AT, gene symbol PI) resides in a cluster of serine protease inhibitor (serpin) genes on chromosome 14q32.1. alpha1AT is highly expressed in the liver and in cultured hepatoma cells. We recently reported the chromatin structure of a >100 kb region around the gene, as defined by DNase I-hypersensitive sites (DHSs) and matrix-attachment regions, in expressing and non-expressing cells. Transfer of human chromosome 14 by microcell fusion from non-expressing fibroblasts to rat hepatoma cells resulted in activation of alpha1AT transcription and chromatin reorganization of the entire region. In the present study, we stably introduced cosmids containing alpha1AT with various amounts of flanking sequence and a linked neo selectable marker into rat hepatoma cells. All single-copy transfectants with >14 kb of 5' flanking sequence expressed wild-type levels of alpha1AT mRNA in a position-independent manner. In contrast, expression of transgenes containing only approximately 1.5-4 kb of flanking sequence was highly variable. Long-term culture of transfectant clones in the absence of selection resulted in gradual loss of neo expression, but expression of the linked alpha1AT gene remained constant. DHS mapping of cosmid transgenes integrated at ectopic sites revealed a hepatoma-specific chromatin structure in each transfectant clone. The implications of these findings are discussed.
Mots-clé
Animals, Carcinoma, Hepatocellular, Chromatin/chemistry, Chromatin/physiology, Chromosomes, Human, Pair 14, Cosmids, Electroporation, Gene Expression, Gene Silencing, Genotype, Hela Cells, Humans, Rats, Transgenes, Tumor Cells, Cultured, alpha 1-Antitrypsin/genetics
Pubmed
Web of science
Création de la notice
11/11/2011 12:10
Dernière modification de la notice
03/03/2018 19:36
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