Global high-throughput screens for cellular function.

There is a critical need for global methods that allow for high-throughput assessment of cellular function for clinical and basic scientists working in both academia and the pharmaceutical industry. These methods typically couple systematic inactivation strategies with high-throughput cell-based assays that facilitate rapid target validation. We present here a survey of these technologies and their applications. We discuss their promise and limitations in addressing the vast number of candidate molecules of disease relevance that are emerging from genomics and proteomics.

[1]  S. Fields,et al.  A biochemical genomics approach for identifying genes by the activity of their products. , 1999, Science.

[2]  A. Saltiel,et al.  Inhibition of MAP Kinase Kinase Blocks the Differentiation of PC-12 Cells Induced by Nerve Growth Factor(*) , 1995, The Journal of Biological Chemistry.

[3]  S. Haggarty,et al.  Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. , 1999, Science.

[4]  J. G. Cory,et al.  Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. , 1991, Cancer communications.

[5]  S. Haggarty,et al.  High-throughput screening of small molecules in miniaturized mammalian cell-based assays involving post-translational modifications. , 1999, Chemistry & biology.

[6]  Stuart L. Schreiber,et al.  Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes , 1991, Cell.

[7]  Paul Schimmel,et al.  M411_3c 107..110 , 2001 .

[8]  J. Shih,et al.  Global analysis of gene expression in invasion by a lung cancer model. , 2001, Cancer research.

[9]  Paul A. Negulescu,et al.  A genome-wide functional assay of signal transduction in living mammalian cells , 1998, Nature Biotechnology.

[10]  P. Zipperlen,et al.  Functional genomic analysis of C. elegans chromosome I by systematic RNA interference , 2000, Nature.

[11]  R F Standaert,et al.  Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin , 1995, Science.

[12]  R. Barstead,et al.  Genome-wide RNAi. , 2001, Current opinion in chemical biology.

[13]  S. Schreiber,et al.  Printing proteins as microarrays for high-throughput function determination. , 2000, Science.

[14]  M. Minden,et al.  Increased sensitivity of acute myeloid leukemias to lovastatin-induced apoptosis: A potential therapeutic approach. , 1999, Blood.

[15]  R. Carthew.  Gene silencing by double-stranded RNA. , 2001, Current opinion in cell biology.

[16]  L. Ilag,et al.  Functional proteomics using chromophore‐ assisted laser inactivation , 2002, Proteomics.

[17]  Christophe Person,et al.  Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells , 1998, Nature.

[18]  J. Hudson,et al.  In vivo gene expression profile analysis of human breast cancer progression. , 1999, Cancer research.

[19]  J. Dowling,et al.  Small molecule developmental screens reveal the logic and timing of vertebrate development. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Y Iwamoto,et al.  A rapid in vitro assay for quantitating the invasive potential of tumor cells. , 1987, Cancer research.

[21]  R. Heim,et al.  Development and Application of a GFP-FRET Intracellular Caspase Assay for Drug Screening , 2000, Journal of biomolecular screening.

[22]  A. Fire,et al.  RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Sebastian A. Leidel,et al.  Functional genomic analysis of cell division in C. elegans using RNAi of genes on chromosome III , 2000, Nature.

[24]  R. Schier,et al.  Fluorophore‐assisted light inactivation: A high‐throughput tool for direct target validation of proteins , 2002, Proteomics.

[25]  B. Eustace,et al.  Fluorophore-assisted light inactivation for multiplex analysis of protein function in cellular processes. , 2003, Methods in enzymology.

[26]  E. Wong,et al.  Study of in situ function of cytoskeletal proteins in lamellipodia and filopodia using microscale chromophore-assisted laser inactivation. , 2000, Methods in enzymology.

[27]  James R. Knight,et al.  A comprehensive analysis of protein–protein interactions in Saccharomyces cerevisiae , 2000, Nature.

[28]  J. Drews.  In Quest of Tomorrow's Medicines , 1999 .

[29]  Sverdrup,et al.  Antisense oligonucleotides: a systematic high-throughput approach to target validation and gene function determination. , 1999, Drug discovery today.

[30]  G. Martin,et al.  Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[32]  Yuji Kohara,et al.  Large-scale analysis of gene function in Caenorhabditis elegans by high-throughput RNAi , 2001, Current Biology.

[33]  S. Asano,et al.  A method to identify cDNAs based on localization of green fluorescent protein fusion products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[34]  J. Mesirov,et al.  Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. , 1999, Science.

[35]  Kei-Hoi Cheung,et al.  Large-scale analysis of the yeast genome by transposon tagging and gene disruption , 1999, Nature.

[36]  D. Jay,et al.  Selective destruction of protein function by chromophore-assisted laser inactivation. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Ronald W. Davis,et al.  Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. , 1999, Science.

[38]  G. Zlokarnik,et al.  Quantitation of transcription and clonal selection of single living cells with beta-lactamase as reporter. , 1998, Science.

[39]  D. J. Clarke,et al.  A topoisomerase II-dependent G2 cycle checkpoint in mammalian cells , 1994, Nature.

[40]  S. Ekker,et al.  Effective targeted gene ‘knockdown’ in zebrafish , 2000, Nature Genetics.

[41]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[42]  Juri Rappsilber,et al.  Mass spectrometry and EST-database searching allows characterization of the multi-protein spliceosome complex , 1998, Nature Genetics.

[43]  S. Cutler,et al.  Random GFP::cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M V Boland,et al.  Automated recognition of patterns characteristic of subcellular structures in fluorescence microscopy images. , 1998, Cytometry.

[45]  J. Summerton.  Morpholino antisense oligomers: the case for an RNase H-independent structural type. , 1999, Biochimica et biophysica acta.

[46]  B. Bochner,et al.  Phenotype microarrays for high-throughput phenotypic testing and assay of gene function. , 2001, Genome research.

[47]  M. Gerstein,et al.  Analysis of yeast protein kinases using protein chips , 2000, Nature Genetics.

[48]  I. Saiki,et al.  A modified and convenient method for assessing tumor cell invasion and migration and its application to screening for inhibitors. , 1997, Biological & pharmaceutical bulletin.

[49]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[50]  Eric S. Lander,et al.  Genomic analysis of metastasis reveals an essential role for RhoC , 2000, Nature.