Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy
Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy
复制标题
通过简单的 TALEN 策略对果蝇基因组进行高效、特异性的修饰
DOI：
10.1016/j.jgg.2012.04.003
复制
发表时间：
2012-05-01
影响因子：
5.9
通讯作者：
Jiao, Renjie
中科院分区：
文献类型：
--
作者：
Liu, Jiyong;Li, Changqing;Jiao, Renjie
技术发展一直是推动生物医学研究取得突破的动力之一。自托马斯·摩根（Thomas Morgan）时代以来，果蝇学家一步一步地开发了强大的遗传工具，用于操纵和功能剖析果蝇基因组，但改进这些技术和开发新技术的空间仍然很大，特别是在生物学家开始以高通量的方式系统地研究包括人类在内的不同模式生物的功能基因组学的今天。在这里，我们首次在果蝇中报道了一种快速、简单和高度特异性的方法，通过改进的转录激活因子样效应核酸酶（TALEN）策略，以非常高的效率修饰果蝇基因组。我们利用了最近发展的“单元组装”策略，组装了两对专门用于修饰黄色基因（在性染色体上）和一个新的常染色体基因的特定TALENs。随后将TALENs mrna注射到果蝇胚胎中。从31.2%的注射F-0可育蝇中，我们检测到涉及黄色基因的遗传修饰。从构建特定的TALENs到检测可继承的修改，整个过程可以在一个月内完成。讨论了talen介导的基因组修饰方法在果蝇中的潜在应用。
Technology development has always been one of the forces driving breakthroughs in biomedical research. Since the time of Thomas Morgan, Drosophilists have, step by step, developed powerful genetic tools for manipulating and functionally dissecting the Drosophila genome, but room for improving these technologies and developing new techniques is still large, especially today as biologists start to study systematically the functional genomics of different model organisms, including humans, in a high-throughput manner. Here, we report, for the first time in Drosophila, a rapid, easy, and highly specific method for modifying the Drosophila genome at a very high efficiency by means of an improved transcription activator-like effector nuclease (TALEN) strategy. We took advantage of the very recently developed "unit assembly" strategy to assemble two pairs of specific TALENs designed to modify the yellow gene (on the sex chromosome) and a novel autosomal gene. The mRNAs of TALENs were subsequently injected into Drosophila embryos. From 31.2% of the injected F-0 fertile flies, we detected inheritable modification involving the yellow gene. The entire process from construction of specific TALENs to detection of inheritable modifications can be accomplished within one month. The potential applications of this TALEN-mediated genome modification method in Drosophila are discussed.