Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases.
Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases.
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DOI：
10.1038/ncomms2020
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发表时间：
2012
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
半变态或不完全变态的昆虫在昆虫遗传学上是相对基础的，并且包括许多害虫。然而，缺乏一个复杂的遗传模型系统，或有针对性的基因操作系统，限制了半代谢物种的研究。在这里，我们使用锌指核酸酶和转录激活因子样效应核酸酶技术，以产生基因敲除半代谢昆虫蟋蟀bimaculatus。将编码锌指核酸酶或转录激活因子样效应物核酸酶的mRNA显微注射到蟋蟀胚胎中后，靶向转基因或内源基因导致序列特异性突变。高达48%的创始人动物后，锌指核酸酶显微注射传输破坏的基因等位基因相比，17%后，显微注射转录激活因子样效应核酸酶。杂合后代使用突变检测测定法选择，所述突变检测测定法使用Surveyor（Cel-I）核酸酶，并且随后的同胞杂交产生纯合敲除蟋蟀。这种方法不依赖于突变体表型或感兴趣的生物体的遗传易处理性，并且可以潜在地应用于使用非转基因策略来管理昆虫害虫。半变态昆虫包括许多害虫，但将靶向突变引入这些物种一直很困难。本文报道了有效的靶向诱变，并产生纯合敲除，在蟋蟀锌指核酸酶或转录激活因子样效应核酸酶的基础上。
Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically relatively basal and comprise many pests. However, the absence of a sophisticated genetic model system, or targeted gene-manipulation system, has limited research on hemimetabolous species. Here we use zinc-finger nuclease and transcription activator-like effector nuclease technologies to produce genetic knockouts in the hemimetabolous insect Gryllus bimaculatus. Following the microinjection of mRNAs encoding zinc-finger nucleases or transcription activator-like effector nucleases into cricket embryos, targeting of a transgene or endogenous gene results in sequence-specific mutations. Up to 48% of founder animals transmit disrupted gene alleles after zinc-finger nucleases microinjection compared with 17% after microinjection of transcription activator-like effector nucleases. Heterozygous offspring is selected using mutation detection assays that use a Surveyor (Cel-I) nuclease, and subsequent sibling crosses create homozygous knockout crickets. This approach is independent from a mutant phenotype or the genetic tractability of the organism of interest and can potentially be applied to manage insect pests using a non-transgenic strategy. Hemimetabolous insects comprise many pests but introducing targeted mutations into these species has been difficult. This paper reports efficient targeted mutagenesis, and the generation of homozygous knockouts, in crickets based on zinc finger nucleases or transcription activator-like effector nucleases.
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影响因子：
3.8
作者：
Takasu, Yoko;Kobayashi, Isao;Zurovec, Michal
通讯作者：
Zurovec, Michal
DOI：
10.1073/pnas.1009424107
发表时间：
2010-08-24
影响因子：
11.1
作者：
Meyer, Melanie;de Angelis, Martin Hrabe;Kuehn, Ralf
通讯作者：
Kuehn, Ralf
影响因子：
46.9
作者：
Szczepek, Michal;Brondani, Vincent;Cathomen, Toni
通讯作者：
Cathomen, Toni
通过简单的 TALEN 策略对果蝇基因组进行高效、特异性的修饰
影响因子：
5.9
作者：
Liu, Jiyong;Li, Changqing;Jiao, Renjie
通讯作者：
Jiao, Renjie
影响因子：
46.9
作者：
Miller, Jeffrey C.;Tan, Siyuan;Rebar, Edward J.
通讯作者：
Rebar, Edward J.