title: Evidence for targeted gene transfer by receptor-mediated endocytosis. Stable expression following insulin-directed entry of NEO into HepG2 cells.
abstract: Evidence is presented for targeted gene delivery to HepG2 cells via the endocytotic pathway under the direction of insulin. Serum albumin was treated with the water-soluble carbodiimide N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide hydrochloride and the resultant positively charged N-acylurea albumin covalently conjugated to insulin by glutaraldehyde cross-linkage. The conjugated protein is shown by nitrocellulose filter binding and gel band shift assays to bind DNA, and by competitive displacement of [125I]insulin to bind to the insulin receptor. When the expression vectors ptkNEO and pAL-8 which incorporate the neo gene were complexed to the conjugate in an in vitro system of transfection, G418 resistant clones developed at frequencies of 2.0-5.5 x 10(-5). Subsequently, a 923bp PstI fragment within the neo sequence was identified by Southern transfer in genomic DNA from transfected cell populations which had been maintained on a G418 regime for 44 days.
authors.affiliation: Department of Biochemistry, University of Durban-Westville, South Africa.
authors.forename: B
authors.initials: B
authors.lastname: Huckett
authors.forename: M
authors.initials: M
authors.lastname: Ariatti
authors.forename: A O
authors.initials: AO
authors.lastname: Hawtrey
mesh_terms: D002460:Cell Line; D004247:DNA / Q000378:metabolism; D004251:DNA Transposable Elements; D004705:Endocytosis; D007328:Insulin / Q000378:metabolism*; D011972:Receptor, Insulin / Q000502:physiology*; D012709:Serum Albumin / Q000378:metabolism; D014162:Transfection
chemical_list: D004251:DNA Transposable Elements; D007328:Insulin; D012709:Serum Albumin; D004247:DNA; D011972:Receptor, Insulin