Retroviral microarray-based platform on nanostructured TiO2 for functional genomics and drug discovery.
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.
Carbone
Roberta
R
Tethis S.r.l., Via Russoli 3, 20151 Milan, Italy.
Giorgetti
Luca
L
Zanardi
Andrea
A
Marangi
Ida
I
Chierici
Elisabetta
E
Bongiorno
Gero
G
Fiorentini
Francesca
F
Faretta
Mario
M
Piseri
Paolo
P
Pelicci
Pier Giuseppe
PG
Milani
Paolo
P
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
D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins; C009495:titanium dioxide; D019809:Streptavidin; D014025:Titanium