Department of Biology, Indiana University, Bloomington, IN, United States of America.
PLoS One. 2018 Apr 12;13(4):e0195977. doi: 10.1371/journal.pone.0195977. eCollection 2018.
The red flour beetle, Tribolium castaneum, is an important model insect and agricultural pest. However, many standard genetic tools are lacking or underdeveloped in this system. Here, we present a set of new reagents to augment existing Tribolium genetic tools. We demonstrate a new GAL4 driver line that employs the promoter of a ribosomal protein gene to drive expression of a UAS responder in the fat body. We also present a novel dual fluorescent reporter that labels cell membranes and nuclei with different fluorophores for the analysis of cellular morphology. This approach also demonstrates the functionality of the viral T2A peptide for bicistronic gene expression in Tribolium. To facilitate classical genetic analysis, we created lines with visible genetic markers by CRISPR-mediated disruption of the yellow and ebony body color loci with a cassette carrying an attP site, enabling future φC31-mediated integration. Together, the reagents presented here will facilitate more robust genetic analysis in Tribolium and serve as a blueprint for the further development of this powerful model's genetic toolkit.
红粉甲虫，Tribolium castaneum，是一种重要的模式昆虫和农业害虫。然而，在这个系统中，许多标准的遗传工具缺乏或发育不全。在这里，我们提出了一组新的试剂来扩充现有的粉甲虫遗传工具。我们展示了一种新的 GAL4 驱动线，该驱动线使用核糖体蛋白基因的启动子在脂肪体中驱动 UAS 应答器的表达。我们还提出了一种新的双荧光报告基因，该报告基因用不同的荧光团标记细胞膜和细胞核，用于分析细胞形态。这种方法还证明了病毒 T2A 肽在粉甲虫中双顺反子基因表达的功能。为了促进经典的遗传分析，我们通过 CRISPR 介导的黄色和乌木体色基因座与携带 attP 位点的盒的破坏，创建了带有可见遗传标记的品系，从而为未来的 φC31 介导的整合提供了便利。总之，这里提出的试剂将促进粉甲虫中更强大的遗传分析，并为进一步开发这种强大模型的遗传工具包提供蓝图。
