Modular system for the construction of zinc-finger libraries and proteins

被引：65

作者：

Gonzalez, Beatriz ^{[1
,2
,3
]}

Schwimmer, Lauren J. ^{[1
,2
,3
]}

Fuller, Roberta P. ^{[1
,2
,3
]}

Ye, Yongjun ^{[1
,2
,3
]}

Asawapornmongkol, Lily ^{[1
,2
,3
]}

Barbas, Carlos F., III ^{[1
,2
,3
]}

机构：

[1] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA

[2] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

[3] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA

来源：

NATURE PROTOCOLS | 2010年 / 5卷 / 04期

基金：

美国国家卫生研究院;

关键词：

ARTIFICIAL TRANSCRIPTION FACTORS; CONTROLLING GENE-EXPRESSION; DNA-BINDING PROTEINS; DOUBLE-STRANDED DNA; GROWTH-FACTOR-A; HUMAN-CELLS; IN-VIVO; HOMOLOGOUS RECOMBINATION; COMBINATORIAL LIBRARIES; PHENOTYPIC ALTERATION;

D O I：

10.1038/nprot.2010.34

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Engineered zinc-finger transcription factors (ZF-TF) are powerful tools to modulate the expression of specific genes. Complex libraries of ZF-TF can be delivered into cells to scan the genome for genes responsible for a particular phenotype or to select the most effective ZF-TF to regulate an individual gene. In both cases, the construction of highly representative and unbiased libraries is critical. In this protocol, we describe a user-friendly ZF technology suitable for the creation of complex libraries and the construction of customized ZF-TFs. The new technology described here simplifies the building of ZF libraries, avoids PCR-introduced bias and ensures equal representation of every module. We also describe the construction of a customized ZF-TF that can be transferred to a number of expression vectors. This protocol can be completed in 9-11 d.

引用

收藏

页码：791 / 810

页数：20

相关论文

共 69 条

[61] Zinc-finger protein-targeted gene regulation: Genomewide single-gene specificity [J].

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 .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (21) :11997-12002

[62] Human immunodeficiency virus type 1 incorporated with fusion proteins consisting of integrase and the designed polydactyl zinc finger protein E2C can bias integration of viral DNA into a predetermined chromosomal region in human cells [J].

Tan, WJ ;

Dong, Z ;

Wilkinson, TA ;

Barbas, CF ;

Chow, SA .

JOURNAL OF VIROLOGY, 2006, 80 (04) :1939-1948

[63] Positive selection of DNA-protein interactions in mammalian cells through phenotypic coupling with retrovirus production [J].

Tschulena, Ulrich ;

Peterson, Kenneth R. ;

Gonzalez, Beatriz ;

Fedosyuk, Halyna ;

Barbas, Carlos F., III .

NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2009, 16 (11) :1195-U10

[64] Highly efficient endogenous human gene correction using designed zinc-finger nucleases [J].

Urnov, FD ;

Miller, JC ;

Lee, YL ;

Beausejour, CM ;

Rock, JM ;

Augustus, S ;

Jamieson, AC ;

Porteus, MH ;

Gregory, PD ;

Holmes, MC .

NATURE, 2005, 435 (7042) :646-651

[65] High-frequency homologous recombination in plants mediated by zinc-finger nucleases [J].

Wright, DA ;

Townsend, JA ;

Winfrey, RJ ;

Irwin, PA ;

Rajagopal, J ;

Lonosky, PM ;

Hall, BD ;

Jondle, MD ;

Voytas, DF .

PLANT JOURNAL, 2005, 44 (04) :693-705

[66] Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly [J].

Wright, David A. ;

Thibodeau-Beganny, Stacey ;

Sander, Jeffry D. ;

Winfrey, Ronnie J. ;

Hirsh, Andrew S. ;

Eichtinger, Magdalena ;

Fu, Fengli ;

Porteus, Matthew H. ;

Dobbs, Drena ;

Voytas, Daniel F. ;

Joung, J. Keith .

NATURE PROTOCOLS, 2006, 1 (03) :1637-1652

[67] Selective inhibition of P-glycoprotein expression in multidrug-resistant tumor cells by a designed transcriptional regulator [J].

Xu, D ;

Ye, DJ ;

Fisher, M ;

Juliano, RL .

JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, 2002, 302 (03) :963-971

[68] Site-directed transposon integration in human cells [J].

Yant, Stephen R. ;

Huang, Yong ;

Akache, Bassel ;

Kay, Mark A. .

NUCLEIC ACIDS RESEARCH, 2007, 35 (07)

[69]

Zhao GC, 2003, ACTA PETROL SIN, V19, P612

← 1 2 3 4 5 6 7 →