Femtosecond infrared laser—an efficient and safe in vivo gene delivery system for prolonged expression
The major advantages of “naked DNA gene therapy” are its simplicity and a low or negligible immune response. Gene delivery by DNA electroporation (EP) involves injection of DNA and the application of a brief electric pulse to enhance cellular permeability. Although EP is an efficient gene transducti...
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| Veröffentlicht in: | Molecular therapy 2003-08, Vol.8 (2), p.342-350 |
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| creator | Zeira, Evelyne Manevitch, Alexandra Khatchatouriants, Artium Pappo, Orit Hyam, Esti Darash-Yahana, Merav Tavor, Einat Honigman, Alik Lewis, Aaron Galun, Eithan |
| description | The major advantages of “naked DNA gene therapy” are its simplicity and a low or negligible immune response. Gene delivery by DNA electroporation (EP) involves injection of DNA and the application of a brief electric pulse to enhance cellular permeability. Although EP is an efficient gene transduction technique in rodents, it requires much higher voltages (>500 V) in larger animals, and hence, in practice it would be hazardous for human patients, as it would cause serious tissue damage. To overcome the obstacles associated with EP-mediated gene delivery in vivo, we developed a new method of gene transduction that uses laser energy. The femtosecond infrared titanium sapphire laser beam was developed specifically for enhancing in vivo gene delivery without risks of tissue damage. System optimization revealed that injection of 10 μg naked DNA into the tibial muscle of mice followed by application of the laser beam for 5 s, focused to 2 mm depth upon an area of 95 × 95 μm2, resulted in the highest intensity and duration of gene expression with no histological or biochemical evidence of muscle damage. We assessed the potential clinical application of LBGT technology by using it to transfer the murine erythropoietin (mEpo) gene into mice. LBGT-mediated mEpo gene delivery resulted in elevated (>22%) hematocrit levels that were sustained for 8 weeks. Gene expression following LBGT was detected for >100 days. Hence, LBGT is a simple, safe, effective, and reproducible method for therapeutic gene delivery with significant clinical potential. |
| doi_str_mv | 10.1016/S1525-0016(03)00184-9 |
| format | Article |
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Gene delivery by DNA electroporation (EP) involves injection of DNA and the application of a brief electric pulse to enhance cellular permeability. Although EP is an efficient gene transduction technique in rodents, it requires much higher voltages (>500 V) in larger animals, and hence, in practice it would be hazardous for human patients, as it would cause serious tissue damage. To overcome the obstacles associated with EP-mediated gene delivery in vivo, we developed a new method of gene transduction that uses laser energy. The femtosecond infrared titanium sapphire laser beam was developed specifically for enhancing in vivo gene delivery without risks of tissue damage. System optimization revealed that injection of 10 μg naked DNA into the tibial muscle of mice followed by application of the laser beam for 5 s, focused to 2 mm depth upon an area of 95 × 95 μm2, resulted in the highest intensity and duration of gene expression with no histological or biochemical evidence of muscle damage. We assessed the potential clinical application of LBGT technology by using it to transfer the murine erythropoietin (mEpo) gene into mice. LBGT-mediated mEpo gene delivery resulted in elevated (>22%) hematocrit levels that were sustained for 8 weeks. Gene expression following LBGT was detected for >100 days. Hence, LBGT is a simple, safe, effective, and reproducible method for therapeutic gene delivery with significant clinical potential.</description><identifier>ISSN: 1525-0016</identifier><identifier>EISSN: 1525-0024</identifier><identifier>DOI: 10.1016/S1525-0016(03)00184-9</identifier><identifier>PMID: 12907157</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; DNA - administration & dosage ; DNA - genetics ; Dose-Response Relationship, Drug ; electroporation ; Electroporation - instrumentation ; Electroporation - methods ; Energy ; Erythropoietin - genetics ; Gene Expression ; gene expression in vivo ; Gene therapy ; Genes, Reporter - genetics ; Genetic engineering ; Genetic Therapy - adverse effects ; Genetic Therapy - instrumentation ; Genetic Therapy - methods ; Infrared Rays ; Lasers ; Methods ; Mice ; naked DNA ; nonviral vectors ; Permeability ; Time Factors ; Transformation, Genetic ; Vectors (Biology)</subject><ispartof>Molecular therapy, 2003-08, Vol.8 (2), p.342-350</ispartof><rights>2003 The American Society of Gene Therapy</rights><rights>Copyright Nature Publishing Group Aug 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c504t-7b1453e5660eefdec600795da128c3d923329074bb43956002dbde005f2cc2a03</citedby><cites>FETCH-LOGICAL-c504t-7b1453e5660eefdec600795da128c3d923329074bb43956002dbde005f2cc2a03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.proquest.com/docview/1792820210?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,64385,64387,64389,72469</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12907157$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zeira, Evelyne</creatorcontrib><creatorcontrib>Manevitch, Alexandra</creatorcontrib><creatorcontrib>Khatchatouriants, Artium</creatorcontrib><creatorcontrib>Pappo, Orit</creatorcontrib><creatorcontrib>Hyam, Esti</creatorcontrib><creatorcontrib>Darash-Yahana, Merav</creatorcontrib><creatorcontrib>Tavor, Einat</creatorcontrib><creatorcontrib>Honigman, Alik</creatorcontrib><creatorcontrib>Lewis, Aaron</creatorcontrib><creatorcontrib>Galun, Eithan</creatorcontrib><title>Femtosecond infrared laser—an efficient and safe in vivo gene delivery system for prolonged expression</title><title>Molecular therapy</title><addtitle>Mol Ther</addtitle><description>The major advantages of “naked DNA gene therapy” are its simplicity and a low or negligible immune response. Gene delivery by DNA electroporation (EP) involves injection of DNA and the application of a brief electric pulse to enhance cellular permeability. Although EP is an efficient gene transduction technique in rodents, it requires much higher voltages (>500 V) in larger animals, and hence, in practice it would be hazardous for human patients, as it would cause serious tissue damage. To overcome the obstacles associated with EP-mediated gene delivery in vivo, we developed a new method of gene transduction that uses laser energy. The femtosecond infrared titanium sapphire laser beam was developed specifically for enhancing in vivo gene delivery without risks of tissue damage. System optimization revealed that injection of 10 μg naked DNA into the tibial muscle of mice followed by application of the laser beam for 5 s, focused to 2 mm depth upon an area of 95 × 95 μm2, resulted in the highest intensity and duration of gene expression with no histological or biochemical evidence of muscle damage. We assessed the potential clinical application of LBGT technology by using it to transfer the murine erythropoietin (mEpo) gene into mice. LBGT-mediated mEpo gene delivery resulted in elevated (>22%) hematocrit levels that were sustained for 8 weeks. Gene expression following LBGT was detected for >100 days. Hence, LBGT is a simple, safe, effective, and reproducible method for therapeutic gene delivery with significant clinical potential.</description><subject>Animals</subject><subject>DNA - administration & dosage</subject><subject>DNA - genetics</subject><subject>Dose-Response Relationship, Drug</subject><subject>electroporation</subject><subject>Electroporation - instrumentation</subject><subject>Electroporation - methods</subject><subject>Energy</subject><subject>Erythropoietin - genetics</subject><subject>Gene Expression</subject><subject>gene expression in vivo</subject><subject>Gene therapy</subject><subject>Genes, Reporter - genetics</subject><subject>Genetic engineering</subject><subject>Genetic Therapy - adverse effects</subject><subject>Genetic Therapy - instrumentation</subject><subject>Genetic Therapy - methods</subject><subject>Infrared Rays</subject><subject>Lasers</subject><subject>Methods</subject><subject>Mice</subject><subject>naked DNA</subject><subject>nonviral vectors</subject><subject>Permeability</subject><subject>Time Factors</subject><subject>Transformation, Genetic</subject><subject>Vectors (Biology)</subject><issn>1525-0016</issn><issn>1525-0024</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkd9KHDEUxkNR6r8-QiUglPZiav5MZjZXItK1gtCL6nXIJCcamUm2yezi3vUhfEKfxIy7KPSmV-cj-Z3vHL6D0GdKvlNCm9PfVDBRkSK_Ev6t1FldyQ9of_vM6p03TZs9dJDzQ1FUyOYj2qNMkpaKdh_dz2EYYwYTg8U-uKQTWNzrDOn575MOGJzzxkMYsS5E1g4Khld-FfEdBMAWer-CtMZ5nUcYsIsJL1LsY7grRvC4SJCzj-EI7TrdZ_i0rYfodv7j5uJndf3r8uri_LoygtRj1Xa0FhxE0xAAZ8E0hLRSWE3ZzHArGefT7nXX1VyK8slsZ4EQ4ZgxTBN-iL5sfMsSf5aQRzX4bKDvdYC4zKrloqaybQp48g_4EJcplN0UbSWbMcLoZCc2lEkx5wROLZIfdForStR0CPV6CDWlrAhXr4dQsvQdb92X3QD2vWubfAHONgCUMFYekspTzAasT2BGZaP_z4gXM72Y0A</recordid><startdate>200308</startdate><enddate>200308</enddate><creator>Zeira, Evelyne</creator><creator>Manevitch, Alexandra</creator><creator>Khatchatouriants, Artium</creator><creator>Pappo, Orit</creator><creator>Hyam, Esti</creator><creator>Darash-Yahana, Merav</creator><creator>Tavor, Einat</creator><creator>Honigman, Alik</creator><creator>Lewis, Aaron</creator><creator>Galun, Eithan</creator><general>Elsevier Inc</general><general>Elsevier Limited</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope></search><sort><creationdate>200308</creationdate><title>Femtosecond infrared laser—an efficient and safe in vivo gene delivery system for prolonged expression</title><author>Zeira, Evelyne ; 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System optimization revealed that injection of 10 μg naked DNA into the tibial muscle of mice followed by application of the laser beam for 5 s, focused to 2 mm depth upon an area of 95 × 95 μm2, resulted in the highest intensity and duration of gene expression with no histological or biochemical evidence of muscle damage. We assessed the potential clinical application of LBGT technology by using it to transfer the murine erythropoietin (mEpo) gene into mice. LBGT-mediated mEpo gene delivery resulted in elevated (>22%) hematocrit levels that were sustained for 8 weeks. Gene expression following LBGT was detected for >100 days. Hence, LBGT is a simple, safe, effective, and reproducible method for therapeutic gene delivery with significant clinical potential.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>12907157</pmid><doi>10.1016/S1525-0016(03)00184-9</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals DNA - administration & dosage DNA - genetics Dose-Response Relationship, Drug electroporation Electroporation - instrumentation Electroporation - methods Energy Erythropoietin - genetics Gene Expression gene expression in vivo Gene therapy Genes, Reporter - genetics Genetic engineering Genetic Therapy - adverse effects Genetic Therapy - instrumentation Genetic Therapy - methods Infrared Rays Lasers Methods Mice naked DNA nonviral vectors Permeability Time Factors Transformation, Genetic Vectors (Biology) |
| title | Femtosecond infrared laser—an efficient and safe in vivo gene delivery system for prolonged expression |
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