Showing papers on "Phenotypic screening published in 2000"
TL;DR: It is demonstrated that small molecules capable of conditional gene product modulation can be identified through developmental screens in zebrafish, and several of the small molecules identified allowed us to dissect the logic of melanocyte and otolith development and to identify critical periods for these events.
Abstract: Much has been learned about vertebrate development by random mutagenesis followed by phenotypic screening and by targeted gene disruption followed by phenotypic analysis in model organisms. Because the timing of many developmental events is critical, it would be useful to have temporal control over modulation of gene function, a luxury frequently not possible with genetic mutants. Here, we demonstrate that small molecules capable of conditional gene product modulation can be identified through developmental screens in zebrafish. We have identified several small molecules that specifically modulate various aspects of vertebrate ontogeny, including development of the central nervous system, the cardiovascular system, the neural crest, and the ear. Several of the small molecules identified allowed us to dissect the logic of melanocyte and otolith development and to identify critical periods for these events. Small molecules identified in this way offer potential to dissect further these and other developmental processes and to identify novel genes involved in vertebrate development.
541 citations
Patent•
22 Mar 2000
TL;DR: In this paper, the use of high-throughput methods for gene targeting, recombination, phenotype screening and biovalidation of drug targets utilizing enhanced homologous recombination (EHR) techniques was described.
Abstract: The invention relates to the use of high-throughput methods for gene targeting, recombination, phenotype screening and biovalidation of drug targets utilizing enhanced homologous recombination (EHR) techniques. These methods utilize robotically driven multichannel pipetters to perform liquid, particle, cell and organism handling, robotically controlled plate and sample handling platforms, magnetic probes and affinity probes to selectively capture nucleic acid hybrids, and thermally regulated plates or blocks for temperature controlled reactions.
25 citations
22 Sep 2000
14 citations
TL;DR: Experiments show that precise gene localization by phenotypic assay of radiation hybrids is practical and was not appreciably impacted by the known instability of such hybrid cells.
Abstract: A high resolution map of the human genome previously has been constructed by using the G3 panel of human/hamster radiation hybrid cell lines and >15,000 unique human genetic markers. By determining whether human DNA sequences are present or absent in each of the hybrids, localization of single genes may routinely be achieved at ≈250-kb resolution. In this paper we have tested whether similarly precise localization might be achieved by phenotypic screening of the hybrids to facilitate positional cloning of unknown genes. We assayed the susceptibility of each of the hybrid cell lines to transduction by retroviral vectors bearing different retroviral envelope proteins that recognize receptors present on human but not on hamster cells. The results for each of the retroviral vectors were informative and allowed precise localization of the receptor genes for the RD114 cat endogenous retrovirus, xenotropic murine leukemia virus, and type C feline leukemia virus. After cloning of the receptors for these retroviruses, we found that standard genotypic mapping by PCR gave results that were nearly identical to those from phenotypic mapping. These experiments show that precise gene localization by phenotypic assay of radiation hybrids is practical and was not appreciably impacted by the known instability of such hybrid cells. This technique should be applicable to many other human genes having discernible phenotypes in hamster cells and, with completion of the human genome project, will allow rapid identification of unknown genes on the basis of phenotype.
11 citations