Beumer Kelly J, Carroll Dana
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84112-5650, United States.
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84112-5650, United States.
Methods. 2014 Jun 15;68(1):29-37. doi: 10.1016/j.ymeth.2013.12.002. Epub 2014 Jan 8.
For a century, Drosophila has been a favored organism for genetic research. However, the array of materials and methods available to the Drosophila worker has expanded dramatically in the last decade. The most common gene targeting tools, zinc finger nucleases, TALENs, and RNA-guided CRISPR/Cas9, have all been adapted for use in Drosophila, both for simple mutagenesis and for gene editing via homologous recombination. For each tool, there exist a number of web sites, design applications, and delivery methods. The successful application of any of these tools also requires an understanding of methods for detecting successful genome modifications. This article provides an overview of the available gene targeting tools and their application in Drosophila. In lieu of simply providing a protocol for gene targeting, we direct the researcher to resources that will allow access to the latest research in this rapidly evolving field.
一个世纪以来,果蝇一直是遗传学研究中备受青睐的生物。然而,在过去十年里,可供果蝇研究人员使用的材料和方法种类大幅增加。最常用的基因靶向工具,锌指核酸酶、转录激活因子样效应物核酸酶(TALENs)以及RNA引导的CRISPR/Cas9,都已被改编用于果蝇研究,既用于简单的诱变,也用于通过同源重组进行基因编辑。对于每种工具,都有许多网站、设计应用程序和递送方法。成功应用这些工具中的任何一种还需要了解检测成功的基因组修饰的方法。本文概述了可用的基因靶向工具及其在果蝇中的应用。我们没有简单地提供基因靶向的方案,而是为研究人员指引了一些资源,以便他们能够获取这个快速发展领域的最新研究成果。