Tightly regulated expression system for eucaryotic cells

Details Tightly regulated expression system for eucaryotic cells Tightly regulated expression system for eucaryotic cells

1993-12-20

1996-12-03

Guzo, David

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4351721, 4352402, 4353201, C12N 1585, C12P 2100

Patent

active

055807526

ABSTRACT:
The invention relates to an expression system which permits control of the levels of protein produced, and optionally provides production of the mature form of the protein directly. The expression systems of the invention include the desired gene downstream of a first DNA which comprises the reverse transcript of an inducible translational regulator and a second DNA which is the reverse transcript of an mRNA capable of self-stabilization. This translated sequence is in turn under the control of a transcriptional promoter which may also be inducible. In a preferred embodiment, the inducible translational regulator is the iron responsive element (IRE) region of ferritin mRNA, and the stabilizing element is also a portion of the ferritin sequence. The expression system is useful, especially, for the production of toxic proteins since protein production can be delayed until desired.

REFERENCES:
patent: 5089397 (1992-02-01), Kushner et al.
Caughman et al. "The Iron-Responsive Element Is the Single Element Responsible for Iron-dependant Translational Regulation of Ferritin Biosynthesis", J. Biol. Chem., vol. 263, No. 35, Dec. 15, 1988, pp. 19048-19052.
Casey et al., Iron Regulation of Transferrin Receptor mRNA Levels Requires ron-responsive Elements and a Rapid Turnover Determinant in the 3' Untranslated Region of te mRNA, EMBO J. 8(12):3693-3699 (1989).
Harrell et al., Ferritin mRNA: Interactions of Iron Regulatory Element with Translational Regulator Protein P-90 and the Effect on Base-pared Flanking Regions, PNAS 88:4166-4170 (1991).
Muller, et al., A Specific mRNA Binding Factor Regulates the Iron-Dependent Stability of Cytoplasmic Transferrin Receptor mRNA, Cell 58:373-382 (1989).
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