Abstract
Small polybasic peptides derived from the transduction domains of certain proteins, such as the third α-helix of the Antennapedia (Antp) homeodomain, can cross the cell membrane through a receptor-independent mechanism. These cell-permeable molecules have been used as 'Trojan horses' to introduce biologically active cargo molecules such as DNA, peptides or proteins into cells. Using these cell-permeable peptides, we have developed an efficient and simple method to increase virally mediated gene delivery and protein expression in vitro and in vivo. Here, we show that cell-permeable peptides increase viral cell entry, improve gene expression at reduced titers of virus and improve efficacy of therapeutically relevant genes in vivo.
This is a preview of subscription content, access via your institution
Relevant articles
Open Access articles citing this article.
-
Application of peptides with an affinity for phospholipid membranes during the automated purification of extracellular vesicles
Scientific Reports Open Access 30 October 2020
-
Combinatorial targeting and discovery of ligand-receptors in organelles of mammalian cells
Nature Communications Open Access 17 April 2012
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
References
Baek, S. & March, K.L. Gene therapy for restenosis: getting nearer the heart of the matter. Circ. Res. 82, 295–305 (1998).
Cullen, B.R. Journey to the center of the cell. Cell 105, 697–700 (2001).
Pizzato, M., Marlow, S.A., Blair, E.D. & Takeuchi, Y. Initial binding of murine leukemia virus particles to cells does not require specific Env-receptor interaction. J. Virol. 73, 8599–8611 (1999).
Sharma, S., Miyanohara, A. & Friedmann, T. Separable mechanisms of attachment and cell uptake during retrovirus infection. J. Virol. 74, 10790–10795 (2000).
Schwarze, S.R., Ho, A., Vocero-Akbani, A. & Dowdy, S.F. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285, 1569–1572 (1999).
Derossi, D., Chassaing, G. & Prochiantz, A. Trojan peptides: the penetratin system for intracellular delivery. Trends Cell Biol. 8, 84–87 (1998).
Bucci, M. et al. In vivo delivery of the caveolin-1 scaffolding domain inhibits nitric oxide synthesis and reduces inflammation. Nat. Med. 6, 1362–1367 (2000).
Derossi, D. et al. Cell internalization of the third helix of the Antennapedia homeodomain is receptor-independent. J. Biol. Chem. 271, 18188–18193 (1996).
Le Doux, J.M., Landazuri, N., Yarmush, M.L. & Morgan, J.R. Complexation of retrovirus with cationic and anionic polymers increases the efficiency of gene transfer. Hum. Gene Ther. 12, 1611–1621 (2001).
Rivard, A. et al. Rescue of diabetes-related impairment of angiogenesis by intramuscular gene therapy with adeno-VEGF. Am. J. Pathol. 154, 355–363 (1999).
Eguchi, A. et al. Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells. J. Biol. Chem. 276, 26204–26210 (2001).
Xia, H., Mao, Q. & Davidson, B.L. The HIV Tat protein transduction domain improves the biodistribution of β-glucuronidase expressed from recombinant viral vectors. Nat. Biotechnol. 19, 640–644 (2001).
Fischer, P.M. et al. Structure-activity relationship of truncated and substituted analogues of the intracellular delivery vector Penetratin. J. Pept. Res. 55, 163–172 (2000).
Luo, Z. et al. Acute modulation of endothelial Akt/PKB activity alters nitric oxide- dependent vasomotor activity in vivo. J. Clin. Invest. 106, 493–499 (2000).
Zheng, L. et al. Cytoprotection of human umbilical vein endothelial cells against apoptosis and CTL-mediated lysis provided by caspase-resistant Bcl-2 without alterations in growth or activation responses. J. Immunol. 164, 4665–4671 (2000).
Scotland, R.S. et al. Functional reconstitution of endothelial nitric oxide synthase reveals the importance of serine 1179 in endothelium-dependent vasomotion. Circ. Res. 90, 904–910 (2002).
Couffinhal, T. et al. Mouse model of angiogenesis. Am. J. Pathol. 152, 1667–1679 (1998).
Acknowledgements
We thank J. Pober and A. Bothwell for the Ret-GFP virus. This work is supported by grants from the US National Institutes of Health (RO1 HL57665, HL61371 and HL64793) to W.C.S. J.P.G. was supported by a fellowship from the Canadian Institutes of Health Research and R.S.S. was funded by a Wellcome Prize Traveling Research Fellowship.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Rights and permissions
About this article
Cite this article
Gratton, JP., Yu, J., Griffith, J. et al. Cell-permeable peptides improve cellular uptake and therapeutic gene delivery of replication-deficient viruses in cells and in vivo. Nat Med 9, 357–362 (2003). https://doi.org/10.1038/nm835
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm835
This article is cited by
-
Application of peptides with an affinity for phospholipid membranes during the automated purification of extracellular vesicles
Scientific Reports (2020)
-
Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides
Molecular Therapy - Methods & Clinical Development (2014)
-
Protein delivery into live cells by incubation with an endosomolytic agent
Nature Methods (2014)
-
Chitosan-mediated non-viral gene delivery with improved serum stability and reduced cytotoxicity
Biotechnology and Bioprocess Engineering (2014)
-
Cationic membrane peptides: atomic-level insight of structure–activity relationships from solid-state NMR
Amino Acids (2013)