Beumer Kelly J, Trautman Jonathan K, Christian Michelle, Dahlem Timothy J, Lake Cathleen M, Hawley R Scott, Grunwald David J, Voytas Daniel F, Carroll Dana
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84112.
G3 (Bethesda). 2013 Oct 3;3(10):1717-25. doi: 10.1534/g3.113.007260.
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-module-to-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.
锌指核酸酶已被证明是在黑腹果蝇和许多其他生物体中进行靶向基因组操作的成功试剂。然而，它们的效用受到新设计显著失败率的限制，这反映了锌指对DNA识别的复杂性。转录激活样效应因子（TALE）DNA结合结构域依赖于一种简单的、一个模块对应一个碱基对的识别密码，并且它们已被非常有效地整合到用于基因组工程的核酸酶（TALENs）中。在本报告中，我们描述了针对果蝇中多个不同基因的TALENs设计，并探讨了TALEN设计的几个参数。TALENs的成功率远高于锌指核酸酶，且诱变频率相当。在几个以前没有此类等位基因的基因中分离到了敲除突变。TALENs是在果蝇中进行靶向基因组操作的有效工具。
