白色基因作为蟋蟀 Gryllus bimaculatus 的转基因标记。

IF 2.1 3区 生物学 Q3 GENETICS & HEREDITY G3: Genes|Genomes|Genetics Pub Date : 2024-10-15 DOI:10.1093/g3journal/jkae235
Emmanuel Gonzalez-Sqalli, Matthieu Caron, Benjamin Loppin
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引用次数: 0

摘要

蟋蟀 Gryllus bimaculatus 是一种新兴的直翅目昆虫模型,被广泛用于各种生物学研究课题。这种半代谢物种似乎与果蝇和其他成熟的全代谢模型具有很强的互补性。为了改进双髻龙的转基因应用,我们设计了一种转化标记基因,其灵感来自于广泛分布的果蝇迷你白+。利用 CRISPR/Cas9,我们首先产生了一个 Gb-white 基因的功能缺失突变等位基因(Gb-w),它在所有发育阶段都表现出白眼。然后我们证明,转基因插入含有 3xP3-Gb-w+ 基因盒的 piggyBac 载体可以挽救眼睛色素沉着。作为一种应用,我们用这种载体产生了表达与 EGFP 融合的中心粒组蛋白 H3 变体(CenH3.1)的 G. bimaculatus 株系,并验证了在蟋蟀中心粒的 EGFP-CenH3.1 检测。最后,我们证明了基于 Minos 的种系转化和使用 ΦC31 整合酶系统的位点特异性质粒插入在 G. bimaculatus 中的功能。
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The white gene as a transgenesis marker for the cricket Gryllus bimaculatus.

The cricket Gryllus bimaculatus is an emerging model insect of the order Orthoptera that is used in a wide variety of biological research themes. This hemimetabolous species appears highly complementary to Drosophila and other well-established holometabolous models. To improve transgenesis applications in G. bimaculatus, we have designed a transformation marker gene inspired from the widespread Drosophila mini-white+. Using CRISPR/Cas9, we first generated a loss-of-function mutant allele of the Gb-white gene (Gb-w), which exhibits a white eye coloration at all developmental stages. We then demonstrate that transgenic insertions of a piggyBac vector containing a 3xP3-Gb-w+ cassette rescue eye pigmentation. As an application, we used this vector to generate G. bimaculatus lines expressing a centromeric histone H3 variant (CenH3.1) fused to EGFP and validated EGFP-CenH3.1 detection at cricket centromeres. Finally, we demonstrate that Minos-based germline transformation and site-specific plasmid insertion with the ΦC31 integrase system function in G. bimaculatus.

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来源期刊
G3: Genes|Genomes|Genetics
G3: Genes|Genomes|Genetics GENETICS & HEREDITY-
CiteScore
5.10
自引率
3.80%
发文量
305
审稿时长
3-8 weeks
期刊介绍: G3: Genes, Genomes, Genetics provides a forum for the publication of high‐quality foundational research, particularly research that generates useful genetic and genomic information such as genome maps, single gene studies, genome‐wide association and QTL studies, as well as genome reports, mutant screens, and advances in methods and technology. The Editorial Board of G3 believes that rapid dissemination of these data is the necessary foundation for analysis that leads to mechanistic insights. G3, published by the Genetics Society of America, meets the critical and growing need of the genetics community for rapid review and publication of important results in all areas of genetics. G3 offers the opportunity to publish the puzzling finding or to present unpublished results that may not have been submitted for review and publication due to a perceived lack of a potential high-impact finding. G3 has earned the DOAJ Seal, which is a mark of certification for open access journals, awarded by DOAJ to journals that achieve a high level of openness, adhere to Best Practice and high publishing standards.
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