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在赤拟谷盗中进行的大规模系统性RNA干扰筛选鉴定出参与昆虫肌肉发育的新基因。

A Large Scale Systemic RNAi Screen in the Red Flour Beetle Identifies Novel Genes Involved in Insect Muscle Development.

作者信息

Schultheis Dorothea, Weißkopf Matthias, Schaub Christoph, Ansari Salim, Dao Van Anh, Grossmann Daniela, Majumdar Upalparna, Hakeemi Muhammad Salim, Troelenberg Nicole, Richter Tobias, Schmitt-Engel Christian, Schwirz Jonas, Ströhlein Nadi, Teuscher Matthias, Bucher Gregor, Frasch Manfred

机构信息

Division of Developmental Biology, Department of Biology, Friedrich-Alexander University of Erlangen-Nürnberg (FAU), 91058 Erlangen, Germany.

Johann-Friedrich-Blumenbach-Institut, GZMB, University of Göttingen, 37077 Göttingen, Germany.

出版信息

G3 (Bethesda). 2019 Apr 9;9(4):1009-1026. doi: 10.1534/g3.118.200995.

Abstract

Although muscle development has been widely studied in there are still many gaps in our knowledge, and it is not known to which extent this knowledge can be transferred to other insects. To help in closing these gaps we participated in a large-scale RNAi screen that used the red flour beetle, , as a screening platform. The effects of systemic RNAi were screened upon double-stranded RNA injections into appropriate muscle-EGFP tester strains. Injections into pupae were followed by the analysis of the late embryonic/early larval muscle patterns, and injections into larvae by the analysis of the adult thoracic muscle patterns. Herein we describe the results of the first-pass screens with pupal and larval injections, which covered ∼8,500 and ∼5,000 genes, respectively, of a total of ∼16,500 genes of the genome. Apart from many genes known from as regulators of muscle development, a collection of genes previously unconnected to muscle development yielded phenotypes in larval body wall and leg muscles as well as in indirect flight muscles. We then present the main candidates from the pupal injection screen that remained after being processed through a series of verification and selection steps. Further, we discuss why distinct though overlapping sets of genes are revealed by the and screening approaches.

摘要

尽管肌肉发育已在[具体昆虫种类未提及]中得到广泛研究,但我们的知识仍存在许多空白,并且尚不清楚这些知识能在多大程度上应用于其他昆虫。为了填补这些空白,我们参与了一项大规模RNA干扰筛选,该筛选以赤拟谷盗[具体昆虫种类未提及]作为筛选平台。在将双链RNA注射到合适的肌肉-增强绿色荧光蛋白(EGFP)测试菌株中后,对系统性RNA干扰的效果进行筛选。向蛹注射后,分析胚胎后期/幼虫早期的肌肉模式,向幼虫注射后,分析成虫胸部肌肉模式。在此,我们描述了对蛹和幼虫注射进行首轮筛选的结果,这些筛选分别覆盖了[具体昆虫种类未提及]基因组总共约16,500个基因中的约8,500个和约5,000个基因。除了许多在[具体昆虫种类未提及]中已知的作为肌肉发育调节因子的基因外,一组先前与肌肉发育无关的基因在幼虫体壁和腿部肌肉以及间接飞行肌肉中产生了表型。然后,我们展示了蛹注射筛选中经过一系列验证和选择步骤后剩下的主要候选基因。此外,我们讨论了为什么[具体昆虫种类未提及]和[具体昆虫种类未提及]的筛选方法会揭示出不同但有重叠的基因集。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5ab5/6469426/d8b0fb3b16a6/1009f1.jpg

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