Human Molecular Genetics, Vol 6, 519-525, Copyright © 1997 by Oxford University Press
RA Montgomery and HC Dietz
This study examines whether the mimicking of selected properties of
naturally occurring antisense RNAs in prokaryotes allows efficient
inhibition of gene expression by in situ-expressed recombinant molecules in
mammalian cells. Prokaryotic regulatory transcripts are expressed at high
levels and have hairpin structures at their termini, features reminiscent
of small nuclear RNAs (snRNAs) which are abundant and stable in the nucleus
of all mammalian cells. A sequence complementary to fibrillin-1 (FBN1)
mRNA, interrupted in its center by a hammerhead ribozyme, was substituted
for the Sm protein binding site between the stem-loop structures of U1
snRNA. Expression of the chimeric antisense RNA resulted in dramatic
inhibition of expression of fibrillin-1 message and protein in stably
transfected cultured cells. The inhibitory effect was localized to the
nucleus. The biological properties of U1 snRNA may provide a widely
applicable vehicle for the in vivo delivery of antisense targeting
sequences.
ARTICLES
Inhibition of fibrillin 1 expression using U1 snRNA as a vehicle for the presentation of antisense targeting sequence
Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
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