Department of Biochemistry, Virginia Tech, 303 Fralin, Blacksburg, VA, 24061, USA.
Fralin Life Science Institute, Virginia Tech, Blacksburg, VA, 24061, USA.
Parasit Vectors. 2018 Dec 24;11(Suppl 2):655. doi: 10.1186/s13071-018-3210-0.
Aedes aegypti is an important mosquito vector that transmits arboviruses that cause devastating diseases including Zika, dengue fever, yellow fever and chikungunya. Improved understanding of gene regulation in the early development of Ae. aegypti will facilitate genetic studies and help the development of novel control strategies of this important disease vector.
In this study, we demonstrated through transgenic assays that the promoter of an endogenous early zygotic gene KLC2 could drive gene expression in the syncytial blastoderm and early cellular blastoderm, which is a stage that the developing germline and the rest of embryo are accessible to genetic manipulation. An unexpected expression of the reporter gene in transgenic male testes was also observed. Further analysis confirmed the expression of the endogenous KLC2 in the testes, which was not detected in the previous RNA sequencing data.
Our finding provided a new promoter element that can be used in future genetic studies and applications in Ae. aegypti. Moreover, our transgenic reporter assays showed that cautions are needed when interpreting RNA sequencing data as transient or tissue-specific transcription may go undetected by RNAseq.
埃及伊蚊是一种重要的蚊媒,可传播导致 Zika、登革热、黄热病和基孔肯雅热等毁灭性疾病的虫媒病毒。更好地了解埃及伊蚊早期发育中的基因调控将有助于遗传研究,并有助于开发这种重要病媒的新控制策略。
在这项研究中,我们通过转基因实验证明,内源性早期合子基因 KLC2 的启动子可驱动合胞胚层和早期细胞胚层中的基因表达,在此阶段,发育中的生殖细胞系和胚胎的其余部分可进行遗传操作。还观察到报告基因在转基因雄性睾丸中的意外表达。进一步的分析证实了内源性 KLC2 在睾丸中的表达,而这在之前的 RNA 测序数据中并未检测到。
我们的发现为未来的遗传研究和在埃及伊蚊中的应用提供了一个新的启动子元件。此外,我们的转基因报告基因检测表明,在解释 RNA 测序数据时需要谨慎,因为瞬时或组织特异性转录可能无法通过 RNAseq 检测到。