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Lookup NU author(s): Dr Kathleen Scougall, Professor James Shaw
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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 0.1μg/ml tetracycline for 48h inhibited pro/insulin secretion to 19-27% and 7-12%, respectively, compared to tetracycline untreated myoblasts. Following a 48h tetracycline incubation (1.0μg/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 72h. 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. © 2003 Elsevier Science (USA). All rights reserved.
Author(s): Scougall KT, Shaw JAM
Publication type: Article
Publication status: Published
Journal: Biochemical and Biophysical Research Communications
Year: 2003
Volume: 304
Issue: 1
Pages: 167-175
ISSN (print): 0006-291X
ISSN (electronic): 1090-2104
Publisher: Academic Press
URL: http://dx.doi.org/10.1016/S0006-291X(03)00553-9
DOI: 10.1016/S0006-291X(03)00553-9
PubMed id: 12705902
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