MersV1, Main, Exploration, bibRecord, 002E64

Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles

Identifieur interne : 002E64 ( Main/Exploration ); précédent : 002E63; suivant : 002E65

Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles

Auteurs : T. L. Fink [États-Unis] ; P. J. Klepcyk [États-Unis] ; S. M. Oette [États-Unis] ; C. R. Gedeon [États-Unis] ; S. L. Hyatt [États-Unis] ; T. H. Kowalczyk [États-Unis] ; R. C. Moen [États-Unis] ; M. J. Cooper [États-Unis]

Source :

Gene therapy : (Basingstoke) [ 0969-7128 ] ; 2006.

RBID : Pascal:06-0326083

Descripteurs français

KwdFr :
- Cellules épithéliales (enzymologie), Expression des gènes, Humains, Lignée cellulaire, Luciferases (analyse), Luciferases (génétique), Microscopie électronique à transmission, Mucoviscidose (), Mucoviscidose (métabolisme), Nanostructures, Nanotechnologie, Plasmides (génétique), Poumon (enzymologie), Protéines à fluorescence verte (analyse), Protéines à fluorescence verte (génétique), Thérapie génétique (), Transfection ().
MESH :
- analyse : Luciferases, Protéines à fluorescence verte.
- enzymologie : Cellules épithéliales, Poumon.
- génétique : Luciferases, Plasmides, Protéines à fluorescence verte.
- métabolisme : Mucoviscidose.
Pascal (Inist)
- Expression des gènes, Humains, Lignée cellulaire, Microscopie électronique à transmission, Mucoviscidose, Nanostructures, Nanotechnologie, Plasmide, Taille, In vivo, Poumon, Thérapie génétique, Transfection, Transfert génétique, DNA, Thérapie génique, Mucoviscidose, Virus.

English descriptors

KwdEn :
- Cell Line, Cystic Fibrosis (metabolism), Cystic Fibrosis (therapy), Cystic fibrosis, DNA, Epithelial Cells (enzymology), Gene Expression, Gene therapy, Genetic Therapy (methods), Genetic transfer, Green Fluorescent Proteins (analysis), Green Fluorescent Proteins (genetics), Humans, In vivo, Luciferases (analysis), Luciferases (genetics), Lung, Lung (enzymology), Microscopy, Electron, Transmission, Nanostructures, Nanotechnology, Plasmid, Plasmids (genetics), Size, Transfection (methods), Virus.
MESH :
- chemical , analysis : Green Fluorescent Proteins, Luciferases.
- enzymology : Epithelial Cells, Lung.
- chemical , genetics : Green Fluorescent Proteins, Luciferases, Plasmids.
- metabolism : Cystic Fibrosis.
- methods : Genetic Therapy, Transfection.
- therapy : Cystic Fibrosis.
- Cell Line, Gene Expression, Humans, Microscopy, Electron, Transmission, Nanostructures, Nanotechnology.

Abstract

Nanoparticles consisting of single molecules of DNA condensed with polyethylene glycol-substituted lysine 30-mers efficiently transfect lung epithelium following intrapulmonary administration. Nanoparticles formulated with lysine polymers having different counterions at the time of DNA mixing have distinct geometric shapes: trifluoroacetate or acetate counterions produce ellipsoids or rods, respectively. Based on intracytoplasmic microinjection studies, nanoparticle ellipsoids having a minimum diameter less than the 25 nm nuclear membrane pore efficiently transfect non-dividing cells. This 25 nm size restriction corresponds to a 5.8 kbp plasmid when compacted into spheroids, whereas the 8-11 nm diameter of rod-like particles is smaller than the nuclear pore diameter. In mice, up to 50% of lung cells are transfected after dosing with a rod-like compacted 6.9kbp lacZ expression plasmid, and correction of the CFTR chloride channel was observed in humans following intranasal administration of a rod-like compacted 8.3 kbp plasmid. To further investigate the potential size and shape limitations of DNA nanoparticles for in vivo lung delivery, reporter gene activity of ellipsoidal and rod-like compacted luciferase plasmids ranging in size between 5.3 and 20.2 kbp was investigated. Equivalent molar reporter gene activities were observed for each formulation, indicating that microinjection size limitations do not apply to the in vivo gene transfer setting.

Affiliations:

Le document en format XML

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<term>Cystic Fibrosis (metabolism)</term>
<term>Cystic Fibrosis (therapy)</term>
<term>Cystic fibrosis</term>
<term>DNA</term>
<term>Epithelial Cells (enzymology)</term>
<term>Gene Expression</term>
<term>Gene therapy</term>
<term>Genetic Therapy (methods)</term>
<term>Genetic transfer</term>
<term>Green Fluorescent Proteins (analysis)</term>
<term>Green Fluorescent Proteins (genetics)</term>
<term>Humans</term>
<term>In vivo</term>
<term>Luciferases (analysis)</term>
<term>Luciferases (genetics)</term>
<term>Lung</term>
<term>Lung (enzymology)</term>
<term>Microscopy, Electron, Transmission</term>
<term>Nanostructures</term>
<term>Nanotechnology</term>
<term>Plasmid</term>
<term>Plasmids (genetics)</term>
<term>Size</term>
<term>Transfection (methods)</term>
<term>Virus</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Cellules épithéliales (enzymologie)</term>
<term>Expression des gènes</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Luciferases (analyse)</term>
<term>Luciferases (génétique)</term>
<term>Microscopie électronique à transmission</term>
<term>Mucoviscidose ()</term>
<term>Mucoviscidose (métabolisme)</term>
<term>Nanostructures</term>
<term>Nanotechnologie</term>
<term>Plasmides (génétique)</term>
<term>Poumon (enzymologie)</term>
<term>Protéines à fluorescence verte (analyse)</term>
<term>Protéines à fluorescence verte (génétique)</term>
<term>Thérapie génétique ()</term>
<term>Transfection ()</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Green Fluorescent Proteins</term>
<term>Luciferases</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Luciferases</term>
<term>Protéines à fluorescence verte</term>
</keywords>
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<term>Poumon</term>
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<term>Lung</term>
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<term>Luciferases</term>
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<term>Plasmides</term>
<term>Protéines à fluorescence verte</term>
</keywords>
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</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Genetic Therapy</term>
<term>Transfection</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Mucoviscidose</term>
</keywords>
<keywords scheme="MESH" qualifier="therapy" xml:lang="en"><term>Cystic Fibrosis</term>
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<term>Gene Expression</term>
<term>Humans</term>
<term>Microscopy, Electron, Transmission</term>
<term>Nanostructures</term>
<term>Nanotechnology</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Expression des gènes</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Microscopie électronique à transmission</term>
<term>Mucoviscidose</term>
<term>Nanostructures</term>
<term>Nanotechnologie</term>
<term>Plasmide</term>
<term>Taille</term>
<term>In vivo</term>
<term>Poumon</term>
<term>Thérapie génétique</term>
<term>Transfection</term>
<term>Transfert génétique</term>
<term>DNA</term>
<term>Thérapie génique</term>
<term>Mucoviscidose</term>
<term>Virus</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Nanoparticles consisting of single molecules of DNA condensed with polyethylene glycol-substituted lysine 30-mers efficiently transfect lung epithelium following intrapulmonary administration. Nanoparticles formulated with lysine polymers having different counterions at the time of DNA mixing have distinct geometric shapes: trifluoroacetate or acetate counterions produce ellipsoids or rods, respectively. Based on intracytoplasmic microinjection studies, nanoparticle ellipsoids having a minimum diameter less than the 25 nm nuclear membrane pore efficiently transfect non-dividing cells. This 25 nm size restriction corresponds to a 5.8 kbp plasmid when compacted into spheroids, whereas the 8-11 nm diameter of rod-like particles is smaller than the nuclear pore diameter. In mice, up to 50% of lung cells are transfected after dosing with a rod-like compacted 6.9kbp lacZ expression plasmid, and correction of the CFTR chloride channel was observed in humans following intranasal administration of a rod-like compacted 8.3 kbp plasmid. To further investigate the potential size and shape limitations of DNA nanoparticles for in vivo lung delivery, reporter gene activity of ellipsoidal and rod-like compacted luciferase plasmids ranging in size between 5.3 and 20.2 kbp was investigated. Equivalent molar reporter gene activities were observed for each formulation, indicating that microinjection size limitations do not apply to the in vivo gene transfer setting.</div>
</front>
</TEI>
<affiliations><list><country><li>États-Unis</li>
</country>
<region><li>Ohio</li>
</region>
</list>
<tree><country name="États-Unis"><region name="Ohio"><name sortKey="Fink, T L" sort="Fink, T L" uniqKey="Fink T" first="T. L." last="Fink">T. L. Fink</name>
</region>
<name sortKey="Cooper, M J" sort="Cooper, M J" uniqKey="Cooper M" first="M. J." last="Cooper">M. J. Cooper</name>
<name sortKey="Fink, T L" sort="Fink, T L" uniqKey="Fink T" first="T. L." last="Fink">T. L. Fink</name>
<name sortKey="Gedeon, C R" sort="Gedeon, C R" uniqKey="Gedeon C" first="C. R." last="Gedeon">C. R. Gedeon</name>
<name sortKey="Hyatt, S L" sort="Hyatt, S L" uniqKey="Hyatt S" first="S. L." last="Hyatt">S. L. Hyatt</name>
<name sortKey="Klepcyk, P J" sort="Klepcyk, P J" uniqKey="Klepcyk P" first="P. J." last="Klepcyk">P. J. Klepcyk</name>
<name sortKey="Klepcyk, P J" sort="Klepcyk, P J" uniqKey="Klepcyk P" first="P. J." last="Klepcyk">P. J. Klepcyk</name>
<name sortKey="Kowalczyk, T H" sort="Kowalczyk, T H" uniqKey="Kowalczyk T" first="T. H." last="Kowalczyk">T. H. Kowalczyk</name>
<name sortKey="Moen, R C" sort="Moen, R C" uniqKey="Moen R" first="R. C." last="Moen">R. C. Moen</name>
<name sortKey="Oette, S M" sort="Oette, S M" uniqKey="Oette S" first="S. M." last="Oette">S. M. Oette</name>
</country>
</tree>
</affiliations>
</record>

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Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles

Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles

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