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Zinc-Finger Protein-Targeted Gene Regulation: Genomewide Single-Gene Specificity

by Snowden, Andrew W. , Campisi, Judith , Qi, Hong , McNamara, Andrew R. , Moore, Michael , Lee, Ya-Li , Berg, Lutz-Peter , Campos, Bradley , Lai, Albert , Spratt, S. Kaye , Jouvenot, Yann , Ullman, Chris , Isalan, Mark , Davalos, Albert , Guschin, Dmitry , Case, Casey C. , Reynolds, Lindsey , Tan, Siyuan , Pabo, Carl O. , Zhang, H. Steven , Howes, Katherine , Gregory, Philip D.

in Base Sequence / Binding Sites - genetics / Biological Sciences / Cell Line / Cell lines / Cells / Checkpoint Kinase 2 / Deoxyribonucleic acid / DNA / DNA - genetics / DNA - metabolism / DNA Damage / Gene expression / Gene Expression Regulation / Gene Expression Regulation, Enzymologic / Genes / Genome, Human / Genomics / HEK293 cells / Humans / Medical treatment / Messenger RNA / Phosphorylation / Promoter Regions, Genetic / Protein Engineering / Protein Kinases - genetics / Protein Kinases - metabolism / Protein Serine-Threonine Kinases / Proteins / Recombinant Proteins - genetics / Recombinant Proteins - metabolism / Repression / Repressor Proteins - genetics / Repressor Proteins - metabolism / RNA, Messenger - genetics / RNA, Messenger - metabolism / Transcription Factors - genetics / Transcription Factors - metabolism / Transfection / Zinc / Zinc Fingers - genetics

2003

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2003

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2003

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Journal Article

Zinc-Finger Protein-Targeted Gene Regulation: Genomewide Single-Gene Specificity

Snowden, Andrew W.,

Campisi, Judith,

Qi, Hong,

McNamara, Andrew R.,

Moore, Michael,

Lee, Ya-Li,

Berg, Lutz-Peter,

Campos, Bradley,

Lai, Albert,

Spratt, S. Kaye,

Jouvenot, Yann,

Ullman, Chris,

Isalan, Mark,

Davalos, Albert,

Guschin, Dmitry,

Case, Casey C.,

Reynolds, Lindsey,

Tan, Siyuan,

Pabo, Carl O.,

Zhang, H. Steven,

Howes, Katherine,

Gregory, Philip D.

2003

Overview

Zinc-finger protein transcription factors (ZFP TFs) can be designed to control the expression of any desired target gene, and thus provide potential therapeutic tools for the study and treatment of disease. Here we report that a ZFP TF can repress target gene expression with single-gene specificity within the human genome. A ZFP TF repressor that binds an 18-bp recognition sequence within the promoter of the endogenous CHK2 gene gives a >10-fold reduction in CHK2 mRNA and protein. This level of repression was sufficient to generate a functional phenotype, as demonstrated by the loss of DNA damage-induced CHK2-dependent p53 phosphorylation. We determined the specificity of repression by using DNA microarrays and found that the ZFP TF repressed a single gene (CHK2) within the monitored genome in two different cell types. These data demonstrate the utility of ZFP TFs as precise tools for target validation, and highlight their potential as clinical therapeutics.

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Publisher

National Academy of Sciences,National Acad Sciences

Subject

Base Sequence

/ Binding Sites - genetics

/ Biological Sciences

/ Cell Line

/ Cell lines

/ Cells

/ Checkpoint Kinase 2