Zinc-finger protein-targeted gene regulation: Genomewide single-gene specificity

被引:130
作者
Tan, SY
Guschin, D
Davalos, A
Lee, YL
Snowden, AW
Jouvenot, Y
Zhang, HS
Howes, K
McNamara, AR
Lai, A
Ullman, C
Reynolds, L
Moore, M
Isalan, M
Berg, LP
Campos, B
Qi, H
Spratt, SK
Case, CC
Pabo, CO
Campisi, J
Gregory, PD
机构
[1] Sangamo Biosci Inc, Point Richmond Tech Ctr 2, Richmond, CA 94804 USA
[2] Univ Calif Berkeley, Lawrence Berkeley Lab, Berkeley, CA 94720 USA
关键词
D O I
10.1073/pnas.2035056100
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:11997 / 12002
页数:6
相关论文
共 40 条
[1]   Questioning the role of checkpoint kinase 2 in the p53 DNA damage response [J].
Ahn, JW ;
Urist, M ;
Prives, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (23) :20480-20489
[2]  
Ahn JY, 2000, CANCER RES, V60, P5934
[3]   THE 65-KDA SUBUNIT OF HUMAN NF-KAPPA-B FUNCTIONS AS A POTENT TRANSCRIPTIONAL ACTIVATOR AND A TARGET FOR V-REL-MEDIATED REPRESSION [J].
BALLARD, DW ;
DIXON, EP ;
PEFFER, NJ ;
BOGERD, H ;
DOERRE, S ;
STEIN, B ;
GREENE, WC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) :1875-1879
[4]   Chk2 kinase - A busy messenger [J].
Bartek, J ;
Falck, J ;
Lukas, J .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2001, 2 (12) :877-886
[5]  
Bartsevich VV, 2000, MOL PHARMACOL, V58, P1
[6]   Positive and negative regulation of endogenous genes by designed transcription factors [J].
Beerli, RR ;
Dreier, B ;
Barbas, CF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (04) :1495-1500
[7]  
Chehab NH, 2000, GENE DEV, V14, P278
[8]   Antisense DNAs as multisite genomic modulators identified by DNA microarray [J].
Cho, YS ;
Kim, MK ;
Cheadle, C ;
Neary, C ;
Becker, KG ;
Cho-Chung, YS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (17) :9819-9823
[9]   siRNAs can function as miRNAs [J].
Doench, JG ;
Petersen, CP ;
Sharp, PA .
GENES & DEVELOPMENT, 2003, 17 (04) :438-442
[10]   A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites [J].
Greisman, HA ;
Pabo, CO .
SCIENCE, 1997, 275 (5300) :657-661