Zinc finger peptides for the regulation of gene expression

被引:170
作者
Klug, A [1 ]
机构
[1] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
关键词
zinc fingers; gene expression; DNA affinity; phage display;
D O I
10.1006/jmbi.1999.3007
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Zinc fingers are small DNA-binding peptide motifs that were discovered in this laboratory. These motifs can be used as modular building blocks for the construction of larger protein domains that recognise and bind to specific DNA sequences. Phage display has been used to create a large library of different zinc fingers from which selections were made for binding to a given DNA sequence. From this database there have been elucidated elements of recognition rules that relate the amino acid sequence of a finger to its preferred DNA binding site. Control of gene expression using designed zinc finger peptides has been demonstrated by the specific inhibition of an oncogene mouse cell Line and also by switching on genes in expression plasmids. These experiments demonstrate that zinc finger DNA-binding domains can be engineered de novo to recognise given DNA sequences. Five to six individual zinc fingers linked together would recognise a DNA sequence 15-18 bp in length, sufficiently long to constitute a rare address in the human genome. By adding functional groups to the engineered :DNA-binding domains, e.g. silencing domains, novel transcription factors can be generated to up- or downregulate expression of a target gene. Among potential applications are the repression of oncogene expression and the disruption of the reproductive cycle of virus infection. (C) 1999 Academic Press.
引用
收藏
页码:215 / 218
页数:4
相关论文
共 19 条
[1]   Promoter-specific activation of gene expression directed by bacteriophage-selected zinc fingers [J].
Choo, Y ;
Castellanos, A ;
GarciaHernandez, B ;
SanchezGarcia, I ;
Klug, A .
JOURNAL OF MOLECULAR BIOLOGY, 1997, 273 (03) :525-532
[2]   SELECTION OF DNA-BINDING SITES FOR ZINC FINGERS USING RATIONALLY RANDOMIZED DNA REVEALS CODED INTERACTIONS [J].
CHOO, Y ;
KLUG, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (23) :11168-11172
[3]   Recognition of DNA methylation by zinc fingers [J].
Choo, Y .
NATURE STRUCTURAL BIOLOGY, 1998, 5 (04) :264-265
[4]   IN-VIVO REPRESSION BY A SITE-SPECIFIC DNA-BINDING PROTEIN DESIGNED AGAINST AN ONCOGENIC SEQUENCE [J].
CHOO, Y ;
SANCHEZGARCIA, I ;
KLUG, A .
NATURE, 1994, 372 (6507) :642-645
[5]   TOWARD A CODE FOR THE INTERACTIONS OF ZINC FINGERS WITH DNA - SELECTION OF RANDOMIZED FINGERS DISPLAYED ON PHAGE [J].
CHOO, Y ;
KLUG, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (23) :11163-11167
[6]   DESIGNING DNA-BINDING PROTEINS ON THE SURFACE OF FILAMENTOUS PHAGE [J].
CHOO, Y ;
KLUG, A .
CURRENT OPINION IN BIOTECHNOLOGY, 1995, 6 (04) :431-436
[7]   LENGTH-ENCODED MULTIPLEX BINDING-SITE DETERMINATION - APPLICATION TO ZINC-FINGER PROTEINS [J].
DESJARLAIS, JR ;
BERG, JM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (23) :11099-11103
[8]   TOWARD RULES RELATING ZINC FINGER PROTEIN SEQUENCES AND DNA-BINDING SITE PREFERENCES [J].
DESJARLAIS, JR ;
BERG, JM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (16) :7345-7349
[9]   THE CRYSTAL-STRUCTURE OF A 2 ZINC-FINGER PEPTIDE REVEALS AN EXTENSION TO THE RULES FOR ZINC-FINGER DNA RECOGNITION [J].
FAIRALL, L ;
SCHWABE, JWR ;
CHAPMAN, L ;
FINCH, JT ;
RHODES, D .
NATURE, 1993, 366 (6454) :483-487
[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