Comparing Zinc Finger Nucleases and Transcription Activator-Like Effector Nucleases for Gene Targeting in Drosophila

被引：49

作者：

Beumer, Kelly J. ^{[1
]}

Trautman, Jonathan K. ^{[1
]}

Christian, Michelle ^{[3
,4
]}

Dahlem, Timothy J. ^{[5
]}

Lake, Cathleen M. ^{[6
]}

Hawley, R. Scott ^{[6
,7
]}

Grunwald, David J. ^{[2
]}

Voytas, Daniel F. ^{[3
,4
]}

Carroll, Dana ^{[1
]}

机构：

[1] Univ Utah, Sch Med, Dept Biochem, Salt Lake City, UT 84112 USA

[2] Univ Utah, Sch Med, Dept Human Genet, Salt Lake City, UT 84112 USA

[3] Univ Minnesota, Dept Genet Cell Biol & Dev, Minneapolis, MN 55455 USA

[4] Univ Minnesota, Ctr Genome Engn, Minneapolis, MN 55455 USA

[5] Univ Utah, Mutat Generat & Detect Core, Hlth Sci Ctr, Salt Lake City, UT 84112 USA

[6] Stowers Inst Med Res, Kansas City, MO 64110 USA

[7] Univ Kansas, Med Ctr, Dept Mol & Integrat Physiol, Kansas City, KS 66160 USA

来源：

G3-GENES GENOMES GENETICS | 2013年 / 3卷 / 10期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

2-HYBRID SELECTION SYSTEM; POOL ENGINEERING OPEN; DNA-SEQUENCES; XANTHINE DEHYDROGENASE; HIGH-EFFICIENCY; HUMAN-CELLS; GENOME; TALENS; DOMAINS; RECOGNITION;

D O I：

10.1534/g3.113.007260

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Zinc-finger nucleases have proven to be successful as reagents for targeted genome manipulation in Drosophila melanogaster and many other organisms. Their utility has been limited, however, by the significant failure rate of new designs, reflecting the complexity of DNA recognition by zinc fingers. Transcription activator-like effector (TALE) DNA-binding domains depend on a simple, one-moduleto- one-base-pair recognition code, and they have been very productively incorporated into nucleases (TALENs) for genome engineering. In this report we describe the design of TALENs for a number of different genes in Drosophila, and we explore several parameters of TALEN design. The rate of success with TALENs was substantially greater than for zinc-finger nucleases, and the frequency of mutagenesis was comparable. Knockout mutations were isolated in several genes in which such alleles were not previously available. TALENs are an effective tool for targeted genome manipulation in Drosophila.

引用

页码：1717 / 1725

页数：9

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