A novel hybrid promoter responsive to pathophysiological and pharmacological regulation.
The aim of this study was to construct a promoter containing DNA motifs for an endogenous transcription factor associated with inflammation along with motifs for pharmacological regulation factors. We demonstrate in transfected cells that expression of a gene of interest is induced by hypoxic conditions or through pharmacological induction, and also show pharmacological repression. In vivo studies utilised electroporation of plasmid to mouse paws, a delivery method shown to be effective by bioluminescence imaging. For gene therapy, the promoter was used to drive expression of IL-1Ra in a paw inflammation model with therapeutic effect observed which was further enhanced when the promoter was additionally induced with a pharmacological activator. One of the most important observations from this study was that promoter induction by hypoxia or inflammation could be prevented by the pharmacological repressor in the absence of doxycycline. These studies demonstrate that hybrid promoters enable pharmacological adjustment to the pathophysiological level of gene expression and, importantly, that they allow termination of gene expression even in the presence of pathophysiological stimuli.
Bone and Joint Research Unit, Barts and The London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, Charterhouse Square, London, UK.
Maria C
MC
Subang
Rewas
R
Fatah
Carly
C
Bright
Patricia
P
Blanco
Mariana
M
Berenstein
Ying
Y
Wu
Osvaldo L
OL
Podhajcer
Paul G
PG
Winyard
Yuti
Y
Chernajovsky
David
D
Gould
D000818:Animals; D015687:Cell Hypoxia; D002460:Cell Line; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000628:therapy*; D053590:Interleukin 1 Receptor Antagonist Protein / Q000235:genetics* / Q000627:therapeutic use; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D059372:Nucleotide Motifs; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects; D013754:Tetracyclines / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection
D053590:Interleukin 1 Receptor Antagonist Protein; D013754:Tetracyclines; D014157:Transcription Factors