ArticleGenome research2001
A streamlined process to phenotypically profile heterologous cDNAs in parallel using yeast cell-based assays.
Article in Genome research, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 30 citations in OpenAlex.
- From beer to breadboards: yeast as a force for biological innovation.Genome biology · 2024Review
- Sentinel interaction mapping - a generic approach for the functional analysis of human disease gene variants using yeast.Disease models & mechanisms · 2020Article
- The functional study of human proteins using humanized yeast.Journal of microbiology (Seoul, Korea) · 2020Review
- Cross-Species Complementation of Nonessential Yeast Genes Establishes Platforms for Testing Inhibitors of Human Proteins.Genetics · 2020Article
- Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants.Genetics · 2015Article
- High-throughput screening system for inhibitors of human Heat Shock Factor 2.Cell stress & chaperones · 2015Article
- Contemporary, yeast-based approaches to understanding human genetic variation.Current opinion in genetics & development · 2013Review
- A survey of yeast genomic assays for drug and target discovery.Pharmacology & therapeutics · 2010Review
- Article
- Assessment of PTEN tumor suppressor activity in nonmammalian models: the year of the yeast.Oncogene · 2008Review
- Discovery of drug-resistant and drug-sensitizing mutations in the oncogenic PI3K isoform p110 alpha.Cancer cell · 2008Article
- Reconstitution of the mammalian PI3K/PTEN/Akt pathway in yeast.The Biochemical journal · 2005Article
Corrections and comments
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Authors and funding
17 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To meet the demands of developing lead drugs for the profusion of human genes being sequenced as part of the human genome project, we developed a high-throughput assay construction method in yeast. A set of optimized techniques allows us to rapidly transfer large numbers of heterologous cDNAs from nonyeast plasmids into yeast expression vectors. These high- or low-copy yeast expression plasmids are then converted quickly into integration-competent vectors for phenotypic profiling of the heterologous gene products. The process was validated first by testing proteins of diverse function, such as p38, poly(ADP-ribose) polymerase-1, and PI 3-kinase, by making active-site mutations and using existing small molecule inhibitors of these proteins. For less well-characterized genes, a novel random mutagenesis scheme was developed that allows a combination selection/screen for mutations that retain full-length expression and yet reverse a growth phenotype in yeast. A broad range of proteins in different functional classes has been profiled, with an average yield for growth interference phenotypes of approximately 30%. The ease of manipulation of the yeast genome affords us the opportunity to approach drug discovery and exploratory biology on a genomic scale and shortens assay development time significantly.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.