title: Phenotypic alteration of eukaryotic cells using randomized libraries of artificial transcription factors.
abstract: We have developed a method in which randomized libraries of zinc finger-containing artificial transcription factors are used to induce phenotypic variations in yeast and mammalian cells. By linking multiple zinc-finger domains together, we constructed more than 100,000 zinc-finger proteins with diverse DNA-binding specificities and fused each of them to either a transcription activation or repression domain. The resulting transcriptional regulatory proteins were expressed individually in cells, and the transfected cells were screened for various phenotypic changes, such as drug resistance, thermotolerance or osmotolerance in yeast, and differentiation in mammalian cells. Genes associated with the selected phenotypes were also identified. Our results show that randomized libraries of artificial transcription factors are useful tools for functional genomics and phenotypic engineering.
authors.affiliation: ToolGen, Inc., 461-6 Jeonmin-Dong, Yuseong-Gu, Daejeon, 305-390, South Korea.
authors.forename: Kyung-Soon
authors.initials: KS
authors.lastname: Park
authors.forename: Dong-ki
authors.initials: DK
authors.lastname: Lee
authors.forename: Horim
authors.initials: H
authors.lastname: Lee
authors.forename: Yangsoon
authors.initials: Y
authors.lastname: Lee
authors.forename: Young-Soon
authors.initials: YS
authors.lastname: Jang
authors.forename: Yong Ha
authors.initials: YH
authors.lastname: Kim
authors.forename: Hyo-Young
authors.initials: HY
authors.lastname: Yang
authors.forename: Seong-il
authors.initials: SI
authors.lastname: Lee
authors.forename: Wongi
authors.initials: W
authors.lastname: Seol
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
authors.forename: Sung-Il
authors.initials: SI
authors.lastname: Lee
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D014644:Genetic Variation; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D032446:Myoblasts / Q000378:metabolism; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D010641:Phenotype; D015202:Protein Engineering / Q000379:methods*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D019151:Peptide Library; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors