Nature Biotechnology21, 269 - 274 (2003)
Published online: 18 February 2003; | doi:10.1038/nbt794
Scanning the human genome with combinatorial transcription factor
libraries
Pilar Blancafort, Laurent Magnenat
& Carlos F. Barbas III
Department of Molecular Biology, The Scripps
Research Institute, 10550 North Torrey Pines Road,
La Jolla, CA 92037.
Correspondence should be addressed to Carlos F. Barbas III carlos@scripps.edu
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.
