Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage
Identifieur interne : 003502 ( Main/Exploration ); précédent : 003501; suivant : 003503Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage
Auteurs : Lorena Urbanelli [Italie] ; Chiara Ronchini [Italie] ; Laura Fontana [Italie] ; Sylvie Menard [Italie] ; Rosaria Orlandi [Italie] ; Paolo Monaci [Italie]Source :
- Journal of Molecular Biology [ 0022-2836 ] ; 2001.
English descriptors
- KwdEn :
- Teeft :
- Assay, Average values, Biol, Capsid, Clone, Elisa, Gene, Gene delivery, Gene delivery vehicles, Gene expression, Gene transduction, Gene transfer, Her2, Independent assays, Lamentous, Lamentous phage, Ligand, Luciferase, Luciferase activity, Luciferase assay, Luciferase expression, Magnetic beads, Mammalian cells, Mewo, Minutes incubation, Monaci, Pbbs, Peptide, Phage, Phage display, Phage particles, Phage vectors, Phagemid, Piii, Plasmid, Pviii, Pviii proteins, Random peptide library, Receptor, Room temperature, Skbr3, Skbr3 cells, Standard deviation, Transduction, Transfection, Triplicate determinations, Unbound, Unbound phage.
Abstract
Abstract: Screening a random peptide library displayed on phage as fusion to the major capsid protein pVIII identified a ligand binding the human epidermal growth factor receptor 2 (HER2) specifically. By mutating the sequence of this ligand, a “secondary” library was generated, whose panning on HER2-positive cells isolated a phage-borne peptide with increased specific binding to HER2 (phage NL1.1). The same peptide recognised HER2 specifically when expressed as an N-terminal fusion to the minor coat protein pIII. Phage NL1.1 was engineered to include a mammalian expression cassette for a reporter gene within its genome. This modified phage transduced HER2-expressing cells with very high specificity (more than 1000-fold that of parental HER2-negative cells) and with an efficiency comparable to that of chemical transfection protocols. The gene delivery process was remarkably fast, requiring less than 15 minutes incubation of phage with target cells to generate detectable levels of gene expression.
Url:
DOI: 10.1006/jmbi.2001.5111
Affiliations:
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P. Angeletti</wicri:orgArea></affiliation></author><author><name sortKey="Ronchini, Chiara" sort="Ronchini, Chiara" uniqKey="Ronchini C" first="Chiara" last="Ronchini">Chiara Ronchini</name><affiliation wicri:level="1"><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Experimental Oncology, Molecular Target Unit, Istituto Nazionale Tumori, Milano</wicri:regionArea><wicri:noRegion>Milano</wicri:noRegion></affiliation></author><author><name sortKey="Fontana, Laura" sort="Fontana, Laura" uniqKey="Fontana L" first="Laura" last="Fontana">Laura Fontana</name><affiliation wicri:level="3"><country>Italie</country><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName><wicri:orgArea>Department of Molecular & Cell Biology, I.R.B.M. P. 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P. Angeletti</wicri:orgArea></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Molecular Biology</title><title level="j" type="abbrev">YJMBI</title><idno type="ISSN">0022-2836</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2001">2001</date><biblScope unit="volume">313</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="965">965</biblScope><biblScope unit="page" to="976">976</biblScope></imprint><idno type="ISSN">0022-2836</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-2836</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>AP</term><term>BSA</term><term>CAR</term><term>CMV</term><term>ECD</term><term>FACS</term><term>FCS</term><term>HER2</term><term>HER2/neu receptor</term><term>HRP</term><term>RLU</term><term>TU</term><term>cancer</term><term>dsDNA</term><term>gene delivery</term><term>mAb</term><term>phage display</term><term>ssDNA</term><term>targeting</term><term>wt</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Assay</term><term>Average values</term><term>Biol</term><term>Capsid</term><term>Clone</term><term>Elisa</term><term>Gene</term><term>Gene delivery</term><term>Gene delivery vehicles</term><term>Gene expression</term><term>Gene transduction</term><term>Gene transfer</term><term>Her2</term><term>Independent assays</term><term>Lamentous</term><term>Lamentous phage</term><term>Ligand</term><term>Luciferase</term><term>Luciferase activity</term><term>Luciferase assay</term><term>Luciferase expression</term><term>Magnetic beads</term><term>Mammalian cells</term><term>Mewo</term><term>Minutes incubation</term><term>Monaci</term><term>Pbbs</term><term>Peptide</term><term>Phage</term><term>Phage display</term><term>Phage particles</term><term>Phage vectors</term><term>Phagemid</term><term>Piii</term><term>Plasmid</term><term>Pviii</term><term>Pviii proteins</term><term>Random peptide library</term><term>Receptor</term><term>Room temperature</term><term>Skbr3</term><term>Skbr3 cells</term><term>Standard deviation</term><term>Transduction</term><term>Transfection</term><term>Triplicate determinations</term><term>Unbound</term><term>Unbound phage</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Screening a random peptide library displayed on phage as fusion to the major capsid protein pVIII identified a ligand binding the human epidermal growth factor receptor 2 (HER2) specifically. By mutating the sequence of this ligand, a “secondary” library was generated, whose panning on HER2-positive cells isolated a phage-borne peptide with increased specific binding to HER2 (phage NL1.1). The same peptide recognised HER2 specifically when expressed as an N-terminal fusion to the minor coat protein pIII. Phage NL1.1 was engineered to include a mammalian expression cassette for a reporter gene within its genome. This modified phage transduced HER2-expressing cells with very high specificity (more than 1000-fold that of parental HER2-negative cells) and with an efficiency comparable to that of chemical transfection protocols. The gene delivery process was remarkably fast, requiring less than 15 minutes incubation of phage with target cells to generate detectable levels of gene expression.</div></front></TEI><affiliations><list><country><li>Italie</li></country><region><li>Latium</li></region><settlement><li>Rome</li></settlement></list><tree><country name="Italie"><region name="Latium"><name sortKey="Urbanelli, Lorena" sort="Urbanelli, Lorena" uniqKey="Urbanelli L" first="Lorena" last="Urbanelli">Lorena Urbanelli</name></region><name sortKey="Fontana, Laura" sort="Fontana, Laura" uniqKey="Fontana L" first="Laura" last="Fontana">Laura Fontana</name><name sortKey="Menard, Sylvie" sort="Menard, Sylvie" uniqKey="Menard S" first="Sylvie" last="Menard">Sylvie Menard</name><name sortKey="Monaci, Paolo" sort="Monaci, Paolo" uniqKey="Monaci P" first="Paolo" last="Monaci">Paolo Monaci</name><name sortKey="Orlandi, Rosaria" sort="Orlandi, Rosaria" uniqKey="Orlandi R" first="Rosaria" last="Orlandi">Rosaria Orlandi</name><name sortKey="Ronchini, Chiara" sort="Ronchini, Chiara" uniqKey="Ronchini C" first="Chiara" last="Ronchini">Chiara Ronchini</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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