Abstract
The peptide nucleic acid (PNA) is a chimeric molecule with the nucleobases connected by peptide bonds. This chimeric nature gives the PNA certain therapeutic advantages over natural antisense nucleic acid molecules. The PNA probes are known for its better and stronger complementation with target nucleic acids. However, cellular delivery of PNA is a major hurdle due to the charge-neutral nature of the PNA. For cellular delivery of PNA, peptide-PNA conjugates are used. This approach may face some practical limitation in terms of PNA antisense activity. In this study, we propose a novel RATH-2 peptide-based non-covalent PNA delivery mechanism. We observed RATH-2 shows a favorable molecular interaction with PNA at 16:1 (peptide:PNA) molar ratio resulting in co-centric nanoparticle formation. With this combination, we could achieve as high as 93% cellular delivery of the PNA. The proposed non-covalent RATH:PNA delivery model showed endocytic entrapment free delivery of PNA. The study further demonstrated the therapeutic application of PNA with in vitro antiviral intervention model. Using RATH-2 non-covalent PNA delivery system, we could inhibit 69.5% viral load. The present study demonstrates a cell-penetrating peptide:PNA interaction can lead to nanoparticle formations that facilitated cellular delivery of PNA.
Key points
• A novel cell-penetrating peptide (RATH-2) was identified for non-covalent delivery of PNA.
• RATH-2 and PNA formed co-centric nanoparticles at appropriate molar combination.
• PNA delivered through the RATH-2 inhibited the viral gene expression and reduced the viral load.
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The experimental generated and analyzed during the present study are available from the corresponding author on reasonable request.
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Acknowledgements
The authors gratefully acknowledge The Director, Indian Veterinary Research Institute (IVRI), for providing the infrastructure and facilities.
Funding
The study was supported by the grant provided by the Department of Biotechnology (DBT), Govt. Of India, New Delhi (Project grant No BT/PR11271/MED/32/81).
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SK, AKT, and MVB conceived and designed the experiment. SK and AKT supervised the experiments. VGJ, KC, and SB performed the experiments. VGJ and SK analyzed data. VGJ, BS, and SK wrote the paper.
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Joshi, V.G., Chindera, K., Bais, M.V. et al. Novel peptide (RATH) mediated delivery of peptide nucleic acids for antiviral interventions. Appl Microbiol Biotechnol 105, 6669–6677 (2021). https://doi.org/10.1007/s00253-021-11502-9
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DOI: https://doi.org/10.1007/s00253-021-11502-9