Antisense Inhibition of δ-Opioid Receptor Gene Function In Vivo by Peptide Nucleic Acids
- 1 April 2000
- journal article
- Published by American Society for Pharmacology & Experimental Therapeutics (ASPET) in Molecular Pharmacology
- Vol. 57 (4), 725-731
- https://doi.org/10.1124/mol.57.4.725
Abstract
Peptide nucleic acids (PNA) are synthetic analogs of DNA that hybridize to complementary oligonucleotide sequences with exceptional affinity and target specificity. The stability of PNA in biological fluids together with the unique hybridization characteristics of these structures suggests that PNA may have considerable potential as antisense agents for experimental use in vivo. To test this hypothesis, we attempted to modulate supraspinal δ-opioid receptor function in rats using PNA sequences designed to be complementary to a region of the rat δ-opioid receptor. Repeated i.c.v. administration of PNA over a period of 5 days significantly inhibited the antinociceptive response and locomotor response to selective δ-opioid receptor agonists. PNA attenuated δ-opioid receptor function in a sequence-specific, target-specific, and reversible manner characteristic of the functional inhibition caused by an antisense mechanism. There were no apparent toxicities arising from the PNA treatment based on the behavior of the animals and inspection of the treated tissues. Saturation binding studies on brain homogenates did not reveal any significant difference in receptor Bmax between treatment groups. However, [35S]guanosine-5′-O-(3-thio)triphosphate binding assays demonstrated a significant decrease in agonist efficacy in homogenates prepared from antisense-treated rats. Taken together, these results demonstrate that peptide nucleic acids are effective antisense agents in vivo and suggest that PNA may be a useful alternative to phosphodiester or phosphorothioate oligonucleotides, or variants thereof, for determination of gene function in vivo.Keywords
This publication has 29 references indexed in Scilit:
- Spinal neurokinin NK1 receptor down-regulation and antinociception: effects of spinal NK1 receptor antisense oligonucleotides and NK1 receptor occupancy.Journal of Neurochemistry, 2002
- Intrastriatal and intraventricular injections of oligodeoxynucleotides in the rat brain: tissue penetration, intracellular distribution and c-fos antisense effectsMolecular Brain Research, 1998
- 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 neuronsNucleic Acids Research, 1998
- Antisense inhibition of gene expression in bacteria by PNA targeted to mRNANature Biotechnology, 1998
- Peptide nucleic acid-targeted mutagenesis of a chromosomal gene in mouse cellsProceedings of the National Academy of Sciences, 1998
- Effects of oligonucleotide length, mismatches and mRNA levels on C-5 propyne-modified antisense potencyNucleic Acids Research, 1996
- An assessment of the antisense properties of RNase H-competent and steric-blocking oligomersNucleic Acids Research, 1995
- NMR Solution Structure of a Peptide Nucleic Acid Complexed with RNAScience, 1994
- PNA hybridizes to complementary oligonucleotides obeying the Watson–Crick hydrogen-bonding rulesNature, 1993
- Cloning of a Delta Opioid Receptor by Functional ExpressionScience, 1992