title: Femtosecond infrared laser-an efficient and safe in vivo gene delivery system for prolonged expression.
abstract: 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 micro g 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 x 95 micro m(2), 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.
authors.affiliation: Goldyne Savad Institute of Gene Therapy, Jerusalem, Israel.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Zeira
authors.forename: Alexandra
authors.initials: A
authors.lastname: Manevitch
authors.forename: Artium
authors.initials: A
authors.lastname: Khatchatouriants
authors.forename: Orit
authors.initials: O
authors.lastname: Pappo
authors.forename: Esti
authors.initials: E
authors.lastname: Hyam
authors.forename: Merav
authors.initials: M
authors.lastname: Darash-Yahana
authors.forename: Einat
authors.initials: E
authors.lastname: Tavor
authors.forename: Alik
authors.initials: A
authors.lastname: Honigman
authors.forename: Aaron
authors.initials: A
authors.lastname: Lewis
authors.forename: Eithan
authors.initials: E
authors.lastname: Galun
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D018274:Electroporation / Q000295:instrumentation / Q000379:methods; D004921:Erythropoietin / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000009:adverse effects / Q000295:instrumentation* / Q000379:methods*; D007259:Infrared Rays; D007834:Lasers; D051379:Mice; D013997:Time Factors; D014170:Transformation, Genetic
chemical_list: D004921:Erythropoietin; D004247:DNA