title: Retroviral microarray-based platform on nanostructured TiO2 for functional genomics and drug discovery.
abstract: Living-cell microarrays are powerful tools for functional genomics and drug discovery. However, despite several attempts to improve this technology, it is still a challenge to obtain microarrays of cells efficiently overexpressing or downregulating specific genes to address complex phenotypes. Here, we present a cell-based microarray for phenotype screening on primary and cancer cells based on the localized reverse infection by retroviruses. Viral vectors are immobilized on a nanostructured titanium dioxide (ns-TiO2) film obtained by depositing a supersonic beam of titania clusters on a glass substrate. We validated the retroviral cell array by overexpression of GFP reporter genes in primary and cancer cells, and by RNA interference of p53 in primary cells by analyzing effects in cell growth. We demonstrate that ns-TiO2 retroviral arrays are an enabling tool for the study of gene function of families of genes for complex phenotypes and for the identification of novel drug targets.
authors.affiliation: Tethis S.r.l., Via Russoli 3, 20151 Milan, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Carbone
authors.forename: Luca
authors.initials: L
authors.lastname: Giorgetti
authors.forename: Andrea
authors.initials: A
authors.lastname: Zanardi
authors.forename: Ida
authors.initials: I
authors.lastname: Marangi
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Chierici
authors.forename: Gero
authors.initials: G
authors.lastname: Bongiorno
authors.forename: Francesca
authors.initials: F
authors.lastname: Fiorentini
authors.forename: Mario
authors.initials: M
authors.lastname: Faretta
authors.forename: Paolo
authors.initials: P
authors.lastname: Piseri
authors.forename: Pier Giuseppe
authors.initials: PG
authors.lastname: Pelicci
authors.forename: Paolo
authors.initials: P
authors.lastname: Milani
mesh_terms: D000818:Animals; D019921:Biotinylation; D015195:Drug Design; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D034622:RNA Interference; D012190:Retroviridae / Q000235:genetics*; D019809:Streptavidin / Q000737:chemistry; D014025:Titanium / Q000737:chemistry*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins; C009495:titanium dioxide; D019809:Streptavidin; D014025:Titanium