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Expression of the blood-clotting factor-VIII cDNA is repressed by a
transcriptional silencer located in its coding region
RC Hoeben, FJ Fallaux, SJ Cramer, DJ van den Wollenberg, H van Ormondt, E Briet and AJ van der Eb
Department of Medical Biochemistry, University of Leiden, The Netherlands.
Hemophilia A is caused by a deficiency of factor-VIII procoagulant (fVIII)
activity. The current treatment by frequent infusions of plasma- derived
fVIII concentrates is very effective but has the risk of transmittance of
blood-borne viruses (human immunodeficiency virus [HIV], hepatitis
viruses). Use of recombinant DNA-derived fVIII as well as gene therapy
could make hemophilia treatment independent of blood- derived products. So
far, the problematic production of the fVIII protein and the low titers of
the fVIII retrovirus stocks have prevented preclinical trials of gene
therapy for hemophilia A in large- animal models. We have initiated a study
of the mechanisms that oppose efficient fVIII synthesis. We have
established that fVIII cDNA contains sequences that dominantly inhibit its
own expression from retroviral as well as from plasmid vectors. The
inhibition is not caused by instability of the fVIII mRNA (t1/2, > or =
6 hours) but rather to repression at the level of transcription. A 305-bp
fragment is identified that is involved in but not sufficient for
repression. This fragment does not overlap the region recently identified
by Lynch et al (Hum Gene Ther 4:259, 1993) as a dominant inhibitor of RNA
accumulation. The repression is mediated by a cellular factor (or factors)
and is independent of the orientation of the element in the transcription
unit, giving the repressor element the hallmarks of a transcriptional
silencer.
Volume 85,
Issue 9,
pp. 2447-2454,
05/01/1995
Copyright © 1995 by The American Society of Hematology
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