Nucleic Acids Research, Vol 26, Issue 21 4910-4916, Copyright © 1998 by Oxford University Press
G Aldrian-Herrada, MG Desarmenien, H Orcel, L Boissin-Agasse, J Mery, J Brugidou and A Rabie
A peptide nucleic acid (PNA) antisense for the AUG translation initiation
region of prepro-oxytocin mRNA was synthesized and coupled to a r
etro-inverso peptide that is rapidly taken up by cells. This bioconjugate
was internalized by cultured cerebral cortex neurons within minutes,
according to the specific property of the vector peptide. The PNA alone
also entered the cells, but more slowly. Cell viability was unaffected when
the PNA concentrations were lower than 10 microM and incubation times less
than for 24 h. Magnocellular neurons from the hypothalamic supraoptic
nucleus, which produce oxytocin and vasopressin, were cultured in
chemically defined medium. Both PNA and vector peptide-PNA depressed the
amounts of the mRNA coding for prepro- oxytocin in these neurons. A
scrambled PNA had no effect and the very cognate prepro-vasopressin mRNA
was not affected. The antisense PNA also depressed the immunocytochemical
signal for prepro-oxytocin in this culture in a dose- and time-dependent
manner. These results show that PNAs driven by the retro-inverso vector
peptide are powerful antisense reagents for use on cells in culture.
ARTICLES
A peptide nucleic acid (PNA) is more rapidly internalized in cultured neurons when coupled to a retro-inverso delivery peptide. The antisense activity depresses the target mRNA and protein in magnocellular oxytocin neurons
CNRS-UPR 9055, Biologie des Neurones Endocrines, CCIPE, 141 rue de la Cardonille, 34094 Montpellier Cedex 5, France.
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