Quantitative analysis of highly parallel transfection in cell microarrays.
As more genomes are sequenced, we are facing the challenge of rapidly unraveling the functions of genes. To that end, cell microarrays have recently been described that transfect thousands of nucleic acids in parallel and can be used to analyze the phenotypic consequences of such perturbations. As many parameters can influence the efficacy of transfection in such a format, we describe some important features in manufacturing cell microarrays that may improve reliability and efficiency of both plasmid DNA and siRNA transfection. We have also developed image analysis software that allows automatic detection of cell clusters, quantification of transfection efficiency and levels of expression/extinction of genes. Along with cell microarrays, this bioinformatic tool should expedite functional exploration of the human genome.
Baghdoyan
Sandrine
S
Service de Génomique Fonctionnelle CEA, 2 Rue Gaston Crémieux CP22, 91057 Evry Cedex, France.
Roupioz
Yoann
Y
Pitaval
Amandine
A
Castel
David
D
Khomyakova
Elena
E
Papine
Alexandre
A
Soussaline
Françoise
F
Gidrol
Xavier
X
D001331:Automation; D001704:Biopolymers; D002460:Cell Line; D002477:Cells / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D005453:Fluorescence; D005780:Gelatin; D020869:Gene Expression Profiling / Q000379:methods; D020868:Gene Silencing; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D015894:Genome, Human; D023281:Genomics / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010641:Phenotype; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012984:Software; D014162:Transfection / Q000379:methods* / Q000592:standards*
D001704:Biopolymers; D008164:Luminescent Proteins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D005780:Gelatin; D004247:DNA