Scanning the human genome with combinatorial transcription factor libraries.
Despite the critical importance of transcription factors in mediating gene regulation, there exists no general, genome-wide tool that uses transcription factors to induce or silence a target gene or select for a particular phenotype. In the strategy described here, we prepared large combinatorial libraries of artificial transcription factors comprising three or six zinc-finger domains, and selected transcription factor-DNA interactions able to upregulate several genes in human cells. Selected transcription factors either induced the expression of an endothelial-specific differentiation marker, VE-cadherin, in non-endothelial cell lines or, when combined with a repression domain, knocked down expression. Potential binding sites for a number of these transcription factors were mapped along the promoter of CDH5, the gene encoding VE-cadherin. Transcription factor libraries represent a useful approach for studying and modulating gene function in cells and potentially in whole organisms.
Blancafort
Pilar
P
Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Magnenat
Laurent
L
Barbas
Carlos F
CF
D002460:Cell Line; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics* / Q000502:physiology; D015894:Genome, Human; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000378:metabolism / Q000502:physiology; D019151:Peptide Library; D040901:Proteomics / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D014162:Transfection / Q000379:methods; D016335:Zinc Fingers / Q000235:genetics
D019151:Peptide Library; D011994:Recombinant Proteins; D014157:Transcription Factors