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Co-transduction of Sleeping Beauty Transposase and Donor Plasmid via a Cell-penetrating Peptide: A simple one-step method
Stockholm University, Faculty of Science, Department of Neurochemistry. University of Tartu, Estonia.
Stockholm University, Faculty of Science, Department of Neurochemistry.
Stockholm University, Faculty of Science, Department of Neurochemistry.
Stockholm University, Faculty of Science, Department of Neurochemistry.ORCID iD: 0000-0003-1443-5892
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2008 (English)In: International journal of peptide research and therapeutics, ISSN 1573-3904, Vol. 14, no 1, 58-63 p.Article in journal (Refereed) Published
Abstract [en]

Transposable elements have emerged as a promising candidate for human non-viral gene-therapy. The Tc1/mariner transposon Sleeping Beauty is to date one of the most efficient transposons in mammals. Sleeping Beauty transposase has so far mostly been delivered to cells via a DNA source. This might cause spontaneous integration of the transposase gene and cause fatal damage to the affected cell. Hence, it would be advantageous to employ a non-genetic source for the transposase. We here show that a novel Cell-penetrating peptide, M918, has the ability to facilitate cellular delivery of both the transposase Sleeping Beauty as a protein and a transposon donor-plasmid carrying an antibiotic resistance gene in vitro. The technique is a simple and straightforward one-step method that might render a safe and efficient delivery platform for Sleeping Beauty mediated gene therapy.

Place, publisher, year, edition, pages
2008. Vol. 14, no 1, 58-63 p.
Keyword [en]
Cell-penetrating peptides, Protein delivery, Transfection, Transposons
National Category
Biological Sciences Chemical Sciences
Research subject
Neurochemistry with Molecular Neurobiology
Identifiers
URN: urn:nbn:se:su:diva-20685DOI: 10.1007/s10989-007-9114-zISI: 000253571800009OAI: oai:DiVA.org:su-20685DiVA: diva2:187211
Available from: 2008-03-04 Created: 2008-03-04 Last updated: 2015-04-21Bibliographically approved
In thesis
1. Cell-penetrating peptides as delivery vectors for oligonucleotides and proteins: Studies on applications and toxicity
Open this publication in new window or tab >>Cell-penetrating peptides as delivery vectors for oligonucleotides and proteins: Studies on applications and toxicity
2007 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Cell-penetrating peptides (CPPs) have for a little bit more than a decade been employed as delivery vectors for a wide range of cargoes, ranging from gold particles to entire plasmids. Although CPP are well studied and utilized in numerous publications, our knowledge about CPP mediated transport is still poor. The articles presented in this thesis all consider different aspects of CPP mediated delivery. The first two papers are evaluating and improving already known techniques. In paper I, standard polyethyleneimine (PEI) transfection is improved by conjugating the CPP TP10 to the cationic polymer. In paper II, the same CPP is employed to deliver a dsDNA decoy oligo, resulting in decreased activity of the transcription factor c-Myc. The third paper is a more general overview of the delivery efficiency of well known CPPs and how the delivered cargo influences the CPP mediated toxicity. The study shows that different CPPs are suitable for different cargos and that toxic side effects depend heavily on the cargo and coupling strategy used. In Paper IV, a novel CPP, M918, is evaluated as a delivery vector for a transposon based non-viral gene therapy system. M918 display simultaneous delivery of a plasmid carrying a selection gene and a transposase into cultured cells. This is the first study where two so vastly different molecules as a cationic protein and an anionic plasmid, are simultaneously transported into cells by a peptide vector. The method might be a first step towards a safe peptide based non-viral gene therapy platform. Taken together, the results presented in this thesis might help to improve already existing techniques, increase our understandings about CPP mediated delivery and, at the same time, develop new CPP based delivery systems.

Place, publisher, year, edition, pages
Stockholm: Institutionen för neurokemi, 2007. 73 p.
National Category
Neurosciences
Research subject
Neurochemistry with Molecular Neurobiology
Identifiers
urn:nbn:se:su:diva-7067 (URN)978-91-7155-499-4 (ISBN)
Public defence
2007-11-02, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 12 A, Stockholm, 13:00 (English)
Opponent
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Available from: 2007-10-12 Created: 2007-09-11 Last updated: 2017-12-01Bibliographically approved

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Järver, PeterTjörnhammar, Marie-LouiseLangel, Ülo
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