Journal
NATURE BIOTECHNOLOGY
Volume 18, Issue 4, Pages 429-432Publisher
NATURE AMERICA INC
DOI: 10.1038/74493
Keywords
temperature-inducible gene expression; Sindbis virus; alphavirus; DNA vector; stable expression; toxic protein
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We present a temperature-regulated, alphavirus replicon-based DNA expression system. The system is regulated by a viral temperature-sensitive RNA-dependent RNA replicase, creating a temperature-dependent RNA amplification loop. Because of this positive feedback, the system exhibits both low background and high inducibility. We observed 700-fold induction in transiently transfected cells, and over 10(4)-fold induction in stably transfected cells. The high stringency of inducibility allowed the generation of stable cell lines expressing a highly toxic protein upon temperature shift. These data suggest that the present expression system could simplify bioprocess engineering strategies, especially in situations where the cloned protein has detrimental effects on host cell metabolism.
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