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  • Title: Tetracycline-regulated secretion of human insulin in transfected primary myoblasts.
    Author: Scougall KT, Shaw JA.
    Journal: Biochem Biophys Res Commun; 2003 Apr 25; 304(1):167-75. PubMed ID: 12705902.
    Abstract:
    A mechanism for safely regulating transgene expression will be necessary for gene therapy approaches to endocrine disorders. In this study, a two-plasmid tetracycline-inducible system was used to regulate expression of human proinsulin (hppI1) and a mutated proinsulin construct (hppI4, allowing cleavage by furin) in primary rat soleus myoblasts. In hppI1 and hppI4 transient transfections, the presence of 0.01 and 0.1 microg/ml tetracycline for 48 h inhibited pro/insulin secretion to 19-27% and 7-12%, respectively, compared to tetracycline untreated myoblasts. Following a 48 h tetracycline incubation (1.0 microg/ml), pro/insulin secretion in hppI1 and hppI4 transfected myoblasts was reduced to <4% of that in cells incubated without tetracycline. Pro/insulin secretion equivalent to that of untreated cells was restored following tetracycline withdrawal and incubation for a further 72 h. Conversion of proinsulin to insulin in transfected myoblasts was <1% for hppI1 and >45% for hppI4. In conclusion, regulated insulin secretion has been achieved in a dose-dependent and reversible manner in primary myoblasts.
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