Positive and Negative Regulation of Endogenous Genes by Designed Transcription Factors

Gene regulation by imposed localization was studied by using designed zinc finger proteins that bind 18-bp DNA sequences in the 5′untranslated regions of the protooncogenes erbB-2 and erbB-3. Transcription factors were generated by fusion of the DNA-binding proteins to repression or activation domai...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2000-02, Vol.97 (4), p.1495-1500
Main Authors: Beerli, Roger R., Dreier, Birgit, Barbas, Carlos F.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies
Biochemistry
Biological Sciences
Cell cycle
Cell Cycle - genetics
Cell Line
Cell lines
DNA
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - pharmacology
erbB-2 gene
erbB-3 gene
Gene expression regulation
Gene Expression Regulation - drug effects
Gene Targeting
Genes
Genes, erbB - genetics
Genes, erbB-2 - genetics
Genes, Reporter
HeLa cells
Humans
Molecular Sequence Data
Proteins
Recombinant Fusion Proteins - pharmacology
Repressor Proteins - pharmacology
Retroviridae
Retroviridae - genetics
Trans-Activators - pharmacology
Transcription factors
Transcription Factors - pharmacology
Transcriptional regulatory elements
Transfection
Zinc Fingers - genetics
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Summary:Gene regulation by imposed localization was studied by using designed zinc finger proteins that bind 18-bp DNA sequences in the 5′untranslated regions of the protooncogenes erbB-2 and erbB-3. Transcription factors were generated by fusion of the DNA-binding proteins to repression or activation domains. When introduced into cells these transcription factors acted as dominant repressors or activators of, respectively, endogenous erbB-2 or erbB-3 gene expression. Significantly, imposed regulation of the two genes was highly specific, despite the fact that the transcription factor binding sites targeted in erbB-2 and erbB-3 share 15 of 18 nucleotides. Regulation of erbB-2 gene expression was observed in cells derived from several species that conserve the DNA target sequence. Repression of erbB-2 in SKBR3 breast cancer cells inhibited cell-cycle progression by inducing a G1accumulation, suggesting the potential of designed transcription factors for cancer gene therapy. These results demonstrate the willful up- and down-regulation of endogenous genes, and provide an additional means to alter biological systems.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.040552697