Correlation between functional and binding activities of designer zinc-finger proteins.
Rapid progress in the ability to develop and utilize zinc-finger proteins with customized sequence specificity have led to their increasing use as tools for modulation of target gene transcription in the post-genomic era. In the present paper, a series of in vitro binding assays and in vivo reporter analyses were used to demonstrate that a zinc-finger protein can effectively specify a base at each position of the target site in vivo and that functional activity of the zinc-finger protein as either a transcriptional repressor or activator is positively correlated with its binding affinity. In addition, this correlation can be extended to artificial engineered zinc-finger proteins. These data suggest that the binding affinity of designer zinc-finger proteins with novel specificity might be a determinant for their ability to regulate transcription of a gene of interest.
Kang
Jong Seok
JS
Department of Cell and Tissue Biology, University of California San Francisco, San Francisco, CA 94143, USA. jong.kang@ucsf.edu
D002460:Cell Line; D051766:Early Growth Response Protein 1 / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008156:Luciferases / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D008156:Luciferases