title: Comparison and optimization of transient transfection methods at human astrocytoma cell line 1321N1.
abstract: Gene delivery to eukaryotic cells is the technique to study the regulation of gene expression. Human astrocytoma cell line 1321N1 could be useful to study G-protein-coupled receptors (GPCRs). Different transient transfection methods, namely calcium phosphate, Lipofectamine, FuGENE, Arrest-In, and microporation (Microporator), were investigated. Results were analyzed by fluorescence-activated cell sorting and fluorescence microscope using green fluorescent protein (GFP) as a reporter gene. To verify the transfection efficiency of these techniques, the expression of human GPR17 gene (hgpr17) was analyzed by transcription quantitative polymerase chain reaction. Microporation resulted in the best method to promote enriched hgpr17 delivery into the human astrocytoma cell line.
authors.affiliation: Medicinal Chemistry Unit, School of Pharmacy, University of Camerino, Camerino, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Marucci
authors.forename: Carmen
authors.initials: C
authors.lastname: Lammi
authors.forename: Michela
authors.initials: M
authors.lastname: Buccioni
authors.forename: Diego
authors.initials: D
authors.lastname: Dal Ben
authors.forename: Catia
authors.initials: C
authors.lastname: Lambertucci
authors.forename: Consuelo
authors.initials: C
authors.lastname: Amantini
authors.forename: Giorgio
authors.initials: G
authors.lastname: Santoni
authors.forename: Meenakshisundaram
authors.initials: M
authors.lastname: Kandhavelu
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Abbracchio
authors.forename: Davide
authors.initials: D
authors.lastname: Lecca
authors.forename: Rosaria
authors.initials: R
authors.lastname: Volpini
authors.forename: Gloria
authors.initials: G
authors.lastname: Cristalli
mesh_terms: D001254:Astrocytoma / Q000378:metabolism; D002130:Calcium Phosphates / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: D002130:Calcium Phosphates; C411955:FuGene; C121670:GPR17 protein, human; D008055:Lipids; C086724:Lipofectamine; D043562:Receptors, G-Protein-Coupled; D049452:Green Fluorescent Proteins; C020243:calcium phosphate