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Toward construction of a self‐sustained clock‐like expression system based on the mammalian circadian clock
Author(s) -
Chilov Dmitri,
Fussenegger Martin
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20143
Subject(s) - circadian clock , transgene , circadian rhythm , microbiology and biotechnology , biology , biopharmaceutical , gene expression , computational biology , neuroscience , genetics , gene
Abstract Despite recent advances in circadian biology, detailed understanding of how a biological pacemaker system is assembled, maintained, and regulated continues to be a significant challenge. We have assembled and characterized a first‐generation, regulatable, self‐sustained clock‐like expression system based on key components of the mammalian circadian clock. The molecular setup of the clock‐like oscillator was reduced to the core set of positive and negative elements common to all known circadian pacemakers. Sophisticated tetracycline‐responsive multi‐cistronic expression integrated with forefront lentiviral transduction tools enabled autoregulated reporter transgene expression in a human cell line. We characterized transgene expression kinetics of an artificial oscillator and showed that its expression profiles could be modulated by a serum shock and administration of regulating tetracycline antibiotics. Design of a generic mammalian clock‐like expression system will offer novel opportunities to study circadian biology and may provide a unique tool for rhythmic expression of desired transgenes fostering advances in biopharmaceutical manufacturing, gene therapy, and tissue engineering. © 2004 Wiley Periodicals, Inc.