Phage display selection of a peptide DNase II inhibitor that enhances gene delivery
Identifieur interne : 002344 ( Main/Exploration ); précédent : 002343; suivant : 002345Phage display selection of a peptide DNase II inhibitor that enhances gene delivery
Auteurs : Jeffrey J. Sperinde [États-Unis] ; Suk-Jung Choi [Corée du Sud] ; Francis C. Szoka Jr [États-Unis]Source :
- The Journal of Gene Medicine [ 1099-498X ] ; 2001-03.
English descriptors
- Teeft :
- Academic press, Active site, Amino acids, Aqueous solution, Assay, Aurintricarboxylic acid, Biochem biophys, Biol chem, Biopharmaceutical sciences, Bovine, Bovine dnase, Cationic, Cationic liposomes, Cell numbers, Charge ratio, Competitive inhibitor, Control peptide, Copyright, Dnase, Dnase activity, Dos, Enhancement, Gene delivery, Gene therapy, Increase transfection, Inhibition, Inhibitor, John wiley sons, Least squares, Liposome, Mdbk, Mdbk cells, Metabolic assay, Modulating transfection, Molecular probes, Monkey kidney, Multiple cell types, Nuclease activity, Nucleic acid therapeutics, Nucleic acids, Pcts1 primer, Peptide, Peptide dnase, Phage, Phage display, Phage library, Plasmid, Plasmid dose, Porcine dnase, Positive effect, Proc natl acad, Sodium acetate, Standard curve, Substrate concentrations, Sulfate, Szoka, Target cell, Total protein, Transfection, Uorescent probe, Various amounts, Wash buffer.
Abstract
Background: Nuclease activity is thought to be a significant barrier to effective gene delivery employing synthetic vectors. In particular, the lysosomal DNase, DNase II, has significant access to plasmid DNA, when the protective condensing agent has been shed. Here, we present the identification of a peptide DNase II inhibitor, enabling enhanced levels of gene delivery. Methods: A DNase II inhibitor was identified by phage display from a cyclic, random 12‐amino acid library. Activity was assayed by inhibition of DNase II degradation of DNA. Transfection enhancement levels were measured over a range of DNA doses with CV‐1 and MDBK cell types using PEI and cationic lipoplexes as vectors. Results: We postulated that a DNase II inhibitor would enhance transfection by enabling a larger fraction of plasmid DNA to traffic through the cell and enter the nucleus. Peptides based on the selected sequence (SLRLLQWFLWAC) [ID2] were shown to inhibit DNase II with an observed KI,app of 0.2–2 µM. Lipoplex‐mediated transfection in vitro was found to be enhanced by ID2‐3 across the entire range of plasmid DNA doses examined (0.10–3.0 µg/mL). Transfection with PEI/DNA complexes was found to be specifically enhanced in the presence of ID2 peptides, with a saturable DNA‐dose curve as would be expected for a competitive inhibitor. Transfection enhancements as high as 270‐fold were found in the presence of ID2‐3. Conclusions: A novel peptide DNase II inhibitor has been used to increase transfection. The level of enhancement was found to be significant in multiple cell types with multiple synthetic vectors. Copyright © 2001 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jgm.165
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001A12
- to stream Istex, to step Curation: 001A12
- to stream Istex, to step Checkpoint: 001170
- to stream Main, to step Merge: 002369
- to stream Main, to step Curation: 002344
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Phage display selection of a peptide DNase II inhibitor that enhances gene delivery</title><author><name sortKey="Sperinde, Jeffrey J" sort="Sperinde, Jeffrey J" uniqKey="Sperinde J" first="Jeffrey J." last="Sperinde">Jeffrey J. Sperinde</name></author><author><name sortKey="Choi, Suk Ung" sort="Choi, Suk Ung" uniqKey="Choi S" first="Suk-Jung" last="Choi">Suk-Jung Choi</name></author><author><name sortKey="Szoka Jr, Francis C" sort="Szoka Jr, Francis C" uniqKey="Szoka Jr F" first="Francis C." last="Szoka Jr">Francis C. Szoka Jr</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:7CC08569E61DE581EA9CB9E3B1BF72FE5B6BC147</idno><date when="2001" year="2001">2001</date><idno type="doi">10.1002/jgm.165</idno><idno type="url">https://api.istex.fr/ark:/67375/WNG-S7MV1JZV-P/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001A12</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001A12</idno><idno type="wicri:Area/Istex/Curation">001A12</idno><idno type="wicri:Area/Istex/Checkpoint">001170</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001170</idno><idno type="wicri:doubleKey">1099-498X:2001:Sperinde J:phage:display:selection</idno><idno type="wicri:Area/Main/Merge">002369</idno><idno type="wicri:Area/Main/Curation">002344</idno><idno type="wicri:Area/Main/Exploration">002344</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">Phage display selection of a peptide DNase II inhibitor that enhances gene delivery</title><author><name sortKey="Sperinde, Jeffrey J" sort="Sperinde, Jeffrey J" uniqKey="Sperinde J" first="Jeffrey J." last="Sperinde">Jeffrey J. Sperinde</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biopharmaceutical Sciences, School of Pharmacy, University of California at San Francisco, San Francisco, CA</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Choi, Suk Ung" sort="Choi, Suk Ung" uniqKey="Choi S" first="Suk-Jung" last="Choi">Suk-Jung Choi</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Chemistry, Kangnung National University, Kangnung</wicri:regionArea><wicri:noRegion>Kangnung</wicri:noRegion></affiliation></author><author><name sortKey="Szoka Jr, Francis C" sort="Szoka Jr, Francis C" uniqKey="Szoka Jr F" first="Francis C." last="Szoka Jr">Francis C. Szoka Jr</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biopharmaceutical Sciences, School of Pharmacy, University of California at San Francisco, San Francisco, CA</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation><affiliation wicri:level="1"><country wicri:rule="url">États-Unis</country></affiliation><affiliation wicri:level="2"><country xml:lang="fr" wicri:curation="lc">États-Unis</country><wicri:regionArea>Correspondence address: Department of Biopharmaceutical Sciences, School of Pharmacy, University of California at San Francisco, 513 Parnassus Avenue, Box 0446, San Francisco, CA 94143‐0446</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">The Journal of Gene Medicine</title><title level="j" type="alt">JOURNAL OF GENE MEDICINE, THE</title><idno type="ISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><imprint><biblScope unit="vol">3</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="101">101</biblScope><biblScope unit="page" to="108">108</biblScope><biblScope unit="page-count">8</biblScope><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2001-03">2001-03</date></imprint><idno type="ISSN">1099-498X</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1099-498X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Academic press</term><term>Active site</term><term>Amino acids</term><term>Aqueous solution</term><term>Assay</term><term>Aurintricarboxylic acid</term><term>Biochem biophys</term><term>Biol chem</term><term>Biopharmaceutical sciences</term><term>Bovine</term><term>Bovine dnase</term><term>Cationic</term><term>Cationic liposomes</term><term>Cell numbers</term><term>Charge ratio</term><term>Competitive inhibitor</term><term>Control peptide</term><term>Copyright</term><term>Dnase</term><term>Dnase activity</term><term>Dos</term><term>Enhancement</term><term>Gene delivery</term><term>Gene therapy</term><term>Increase transfection</term><term>Inhibition</term><term>Inhibitor</term><term>John wiley sons</term><term>Least squares</term><term>Liposome</term><term>Mdbk</term><term>Mdbk cells</term><term>Metabolic assay</term><term>Modulating transfection</term><term>Molecular probes</term><term>Monkey kidney</term><term>Multiple cell types</term><term>Nuclease activity</term><term>Nucleic acid therapeutics</term><term>Nucleic acids</term><term>Pcts1 primer</term><term>Peptide</term><term>Peptide dnase</term><term>Phage</term><term>Phage display</term><term>Phage library</term><term>Plasmid</term><term>Plasmid dose</term><term>Porcine dnase</term><term>Positive effect</term><term>Proc natl acad</term><term>Sodium acetate</term><term>Standard curve</term><term>Substrate concentrations</term><term>Sulfate</term><term>Szoka</term><term>Target cell</term><term>Total protein</term><term>Transfection</term><term>Uorescent probe</term><term>Various amounts</term><term>Wash buffer</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Background: Nuclease activity is thought to be a significant barrier to effective gene delivery employing synthetic vectors. In particular, the lysosomal DNase, DNase II, has significant access to plasmid DNA, when the protective condensing agent has been shed. Here, we present the identification of a peptide DNase II inhibitor, enabling enhanced levels of gene delivery. Methods: A DNase II inhibitor was identified by phage display from a cyclic, random 12‐amino acid library. Activity was assayed by inhibition of DNase II degradation of DNA. Transfection enhancement levels were measured over a range of DNA doses with CV‐1 and MDBK cell types using PEI and cationic lipoplexes as vectors. Results: We postulated that a DNase II inhibitor would enhance transfection by enabling a larger fraction of plasmid DNA to traffic through the cell and enter the nucleus. Peptides based on the selected sequence (SLRLLQWFLWAC) [ID2] were shown to inhibit DNase II with an observed KI,app of 0.2–2 µM. Lipoplex‐mediated transfection in vitro was found to be enhanced by ID2‐3 across the entire range of plasmid DNA doses examined (0.10–3.0 µg/mL). Transfection with PEI/DNA complexes was found to be specifically enhanced in the presence of ID2 peptides, with a saturable DNA‐dose curve as would be expected for a competitive inhibitor. Transfection enhancements as high as 270‐fold were found in the presence of ID2‐3. Conclusions: A novel peptide DNase II inhibitor has been used to increase transfection. The level of enhancement was found to be significant in multiple cell types with multiple synthetic vectors. Copyright © 2001 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><country name="États-Unis"><region name="Californie"><name sortKey="Sperinde, Jeffrey J" sort="Sperinde, Jeffrey J" uniqKey="Sperinde J" first="Jeffrey J." last="Sperinde">Jeffrey J. Sperinde</name></region><name sortKey="Szoka Jr, Francis C" sort="Szoka Jr, Francis C" uniqKey="Szoka Jr F" first="Francis C." last="Szoka Jr">Francis C. Szoka Jr</name><name sortKey="Szoka Jr, Francis C" sort="Szoka Jr, Francis C" uniqKey="Szoka Jr F" first="Francis C." last="Szoka Jr">Francis C. Szoka Jr</name><name sortKey="Szoka Jr, Francis C" sort="Szoka Jr, Francis C" uniqKey="Szoka Jr F" first="Francis C." last="Szoka Jr">Francis C. Szoka Jr</name></country><country name="Corée du Sud"><noRegion><name sortKey="Choi, Suk Ung" sort="Choi, Suk Ung" uniqKey="Choi S" first="Suk-Jung" last="Choi">Suk-Jung Choi</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002344 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002344 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= ChloroquineV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:7CC08569E61DE581EA9CB9E3B1BF72FE5B6BC147
|texte= Phage display selection of a peptide DNase II inhibitor that enhances gene delivery
}}
| This area was generated with Dilib version V0.6.33. |  |