Regulated expression of the interferon-beta gene in mice.
A single plasmid regulated expression vector based upon a mifepristone-inducible two plasmid system, termed pBRES, has been constructed and tested in mice using murine interferon-b (mIFNb) as the transgene. The expression of mIFNb in the circulation was followed by measuring the systemic induction of IP-10, a validated biomarker for mIFNb in mice. Long-term, inducible expression of mIFNb was demonstrated following a single intramuscular (i.m.) injection of the pBRES mIFNb plasmid vector into the hind limb of mice. Induction of mIFNb expression was achieved by administration of the small molecule inducer, mifepristone (MFP). Plasmid DNA and mIFNb mRNA levels in the injected muscles correlated with mIFNb expression as monitored by IP-10 over a 3-month time period. Renewable transgene expression was achieved following repeat administration of the plasmid at 3 months following the first plasmid injection. A dose-dependent increase in expression was demonstrated by varying the amount of injected plasmid or the amount of the inducer administered to the mice. Finally, the pBRES plasmid expressing mIFNb under control of the inducer, MFP, was shown to be efficacious in a murine model of experimental allergic encephalomyelitis, supporting the feasibility of gene-based therapeutic approaches for treating diseases such as multiple sclerosis.
Harkins
R N
RN
Department of Gene Technologies, Berlex Biosciences, Richmond, CA, USA. rick.harkins@bayer.com
Szymanski
P
P
Petry
H
H
Brooks
A
A
Qian
H S
HS
Schaefer
C
C
Kretschmer
P J
PJ
Orme
A
A
Wang
P
P
Rubanyi
G M
GM
Hermiston
T W
TW
D000818:Animals; D015415:Biomarkers / Q000097:blood; D054357:Chemokine CXCL10 / Q000032:analysis; D018450:Disease Progression; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000628:therapy*; D005260:Female; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D007273:Injections, Intramuscular; D016899:Interferon-beta / Q000097:blood / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D015735:Mifepristone / Q000008:administration & dosage; D009103:Multiple Sclerosis / Q000628:therapy; D010957:Plasmids / Q000008:administration & dosage* / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes
D015415:Biomarkers; D054357:Chemokine CXCL10; D012333:RNA, Messenger; D015735:Mifepristone; D016899:Interferon-beta