OPTIMIZATION OF CATIONIC LIPID TECHNIQUES FOR THE DIRECT DELIVERY AND EXPRESSION OF GENES IN VIVO

THE PURPOSE OF THIS PROJECT IS TO DEVELOP AN ASSAY SYSTEM WITH HIGH THROUGHPUT THAT IS CAPABLE OF SCREENING THE EFFICIENCY OF POLYNUCLEOTIDE DELIVERY SYSTEMS UNDER VARIOUS CONDITIONS IN TISSUE CULTURE. SEVERAL NOVEL CATIONIC LIPID ANALOGS AND A VARIETY OF LIPID VEHICLE COMPOSITIONS WILL BE QUANTITATIVELY SCREENED FOR FUNCTIONAL DELIVERY AND EXPRESSION OF A REPORTER GENE PRODUCT IN THE PRESENCE AND ABSENCE OF SERUM. THE AIM WILL BE TO IDENTIFY CANDIDATE FORMULATIONS THAT LEAD TO IMPROVED DELIVERY AND EXPRESSION OF GENES IN VIVO. IN ADDITION TO IDENTIFYING CANDIDATE FORMULATIONS FOR IN VIVO DELIVERY OF POLYNUCLEOTIDES, THIS PROJECT WILL RESULT IN INTERIM PRODUCTS WITH SIGNIFICANT SCIENTIFIC AND COMMERCIAL POTENTIAL AS MOLEUCLAR BIOLOGY REAGENTS. THESE PRODUCTS WILL IMPROVE ON THE EXISTING LIPOFECTIN TRANSFECTION REGENT. THE IMPROVED PRODUCT THAT EMERGES FROM THIS PROJECT WILL RESULT IN GREATER IN VITRO TRANSFECTION ACTIVITY AND REDUCEDCELLULAR TOXICITY. IN ADDITION, THE PRODUCT WILL BE TESTED FOR ACTIVITY ON EMBRYONIC STEM CELLS WHICH, IF ACTIVE, COULDFACILITATE THE STUDY OF TRANSGENIC ANIMALS PRODUCED BY HOMOLOGOUS RECOMBINATION USING CATIONIC LIPID-MEDIATED IN VITRO TRANSFECTION INSTEAD OF ELECTROPORATION.