GeneKnockout by Targeted Mutagenesis in a Hemimetabolous Insect, the Two-Spotted Cricket Gryllus bimaculatus, using TALENs.
GeneKnockout by Targeted Mutagenesis in a Hemimetabolous Insect, the Two-Spotted Cricket Gryllus bimaculatus, using TALENs.
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DOI：
10.1007/978-1-4939-2932-0_12
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发表时间：
2016
影响因子：
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通讯作者：
Takahito Watanabe;S. Noji;T. Mito
中科院分区：
文献类型：
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作者：
Takahito Watanabe;S. Noji;T. Mito
半变态或不完全变态的昆虫是昆虫发生学的基础。这些昆虫包括许多有害物种。蟋蟀（Gryllus bimaculatus）是半代谢昆虫的一种新兴模型，其基础是基于RNA干扰（RNAi）的基因功能分析和转基因技术的成功。利用这种基因组编辑技术将极大地促进功能基因组学研究。使用转录激活因子样效应核酸酶（TALEN）的基因组编辑已被证明是用于在各种物种中进行位点特异性基因组操作的有效方法。TALEN是能够诱导DNA双链断裂进入特定靶序列的人工核酸酶。在这里，我们描述了一种基于TALEN的基因敲除方案。本发明提供了一种用于产生纯合敲除生物体的方法，所述方法包括通过突变检测测定的突变体选择方案。
Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically basal. These insects include many deleterious species. The cricket,Gryllus bimaculatus, is an emerging model for hemimetabolous insects, based on the success of RNA interference (RNAi)-based gene-functional analyses and transgenic technology. Taking advantage of genome-editing technologies in this species would greatly promote functional genomics studies. Genome editing using transcription activator-like effector nucleases (TALENs) has proven to be an effective method for site-specific genome manipulation in various species. TALENs are artificial nucleases that are capable of inducing DNA double-strand breaks into specified target sequences. Here, we describe a protocol for TALEN-based gene knockout inG. bimaculatus, including a mutant selection scheme via mutation detection assays, for generating homozygous knockout organisms.