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Lipid-based Transfe...
Lipid-based Transfection Reagents Exhibit Cryo-induced Increase in Transfection Efficiency
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- Sork, Helena (author)
- Karolinska Institutet
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- Nordin, Joel Z. (author)
- Karolinska Institutet
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Turunen, Janne J. (author)
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- Wiklander, Oscar P. B. (author)
- Karolinska Institutet
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Bestas, Burcu (author)
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- Zaghloul, Eman M. (author)
- Karolinska Institutet
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Margus, Helerin (author)
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Padari, Kärt (author)
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- Duru, Adil D. (author)
- Karolinska Institutet
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- Corso, Giulia (author)
- Karolinska Institutet
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- Bost, Jeremy (author)
- Karolinska Institutet
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Vader, Pieter (author)
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Pooga, Margus (author)
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- Smith, C. I. Edvard (author)
- Karolinska Institutet
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Wood, Matthew J. A. (author)
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Schiffelers, Raymond M. (author)
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- Hällbrink, Mattias (author)
- Stockholms universitet,Institutionen för neurokemi
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- Andaloussi, Samir E. L. (author)
- Karolinska Institutet
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(creator_code:org_t)
- Elsevier BV, 2016
- 2016
- English.
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In: Molecular Therapy Nucleic Acids. - : Elsevier BV. - 2162-2531. ; 5
- Related links:
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https://doi.org/10.1...
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http://www.cell.com/...
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https://urn.kb.se/re...
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Abstract
Subject headings
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- The advantages of lipid-based transfection reagents have permitted their widespread use in molecular biology and gene therapy. This study outlines the effect of cryo-manipulation of a cationic lipid-based formulation, Lipofectamine 2000, which, after being frozen and thawed, showed orders of magnitude higher plasmid delivery efficiency throughout eight different cell lines, without compromising cell viability. Increased transfection efficiency with the freeze-thawed reagent was also seen with 2'-O-methyl phosphorothioate oligonucleotide delivery and in a splice-correction assay. Most importantly, a log-scale improvement in gene delivery using the freeze-thawed reagent was seen in vivo. Using three different methods, we detected considerable differences in the polydispersity of the different nucleic acid complexes as well as observed a clear difference in their surface spreading and sedimentation, with the freeze-thawed ones displaying substantially higher rate of dispersion and deposition on the glass surface. This hitherto overlooked elevated potency of the freeze-thawed reagent facilitates the targeting of hard-to-transfect cells, accomplishes higher transfection rates, and decreases the overall amount of reagent needed for delivery. Additionally, as we also saw a slight increase in plasmid delivery using other freeze-thawed transfection reagents, we postulate that freeze-thawing might prove to be useful for an even wider variety of transfection reagents.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Keyword
- freezing
- Lipofectamine 2000
- lipofection
- lipoplex
- transfection
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Sork, Helena
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Nordin, Joel Z.
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Turunen, Janne J ...
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Wiklander, Oscar ...
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Bestas, Burcu
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Zaghloul, Eman M ...
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show more...
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Margus, Helerin
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Padari, Kärt
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Duru, Adil D.
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Corso, Giulia
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Bost, Jeremy
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Vader, Pieter
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Pooga, Margus
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Smith, C. I. Edv ...
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Wood, Matthew J. ...
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Schiffelers, Ray ...
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Hällbrink, Matti ...
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Andaloussi, Sami ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
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and Medical Biotechn ...
- Articles in the publication
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Molecular Therap ...
- By the university
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Stockholm University
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Karolinska Institutet