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Peptide conjugates for chromosomal gene targeting by triplex-forming oligonucleotides
Faye A Rogers1, Muthiah Manoharan, Peter Rabinovitch
1Department of Therapeutic Radiology, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 068520, USA.
Nucleic Acids Research
|December 17, 2004
Summary
Cell-penetrating peptides enhance the delivery and activity of triplex-forming oligonucleotides (TFOs). Conjugating TFOs to the Antp peptide significantly increased gene-targeted mutagenesis, improving their potential as antigene therapeutics.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Triplex-forming oligonucleotides (TFOs) are DNA-binding molecules with potential for gene expression modulation.
- Cellular uptake limitations hinder the biological activity of TFOs as antigene compounds.
Purpose of the Study:
- To investigate the effect of cell-penetrating peptides on TFO biological activity.
- To assess TFO-peptide conjugates for gene-targeted mutagenesis.
Main Methods:
- TFOs were conjugated to the Antp cell-penetrating peptide.
- Gene-targeted mutagenesis assays were performed comparing TFO-peptide conjugates with unmodified TFOs.
- Confocal microscopy was used to evaluate cellular uptake of labeled TFOs.
Main Results:
- TFOs covalently linked to Antp showed a 20-fold increase in mutation frequency compared to unmodified TFOs.
- Antp alone or linked to control oligonucleotides did not increase mutation frequency.
- TFO-peptide conjugates demonstrated dose-responsive, gene-specific mutagenesis and increased cellular uptake.
Conclusions:
- Peptide conjugation, specifically with Antp, significantly enhances the cellular uptake and biological activity of TFOs.
- This strategy holds promise for improving the intranuclear delivery of DNA-binding reagents for therapeutic applications.