Abstract:
The Masculinizer gene, Masc, encodes a lepidopteran-specific novel CCCH-type zinc finger protein, which controls sex determination and dosage compensation in Bombyx mori. Considering the potential application of it in pest control, it is necessary to investigate the function of Masc gene in Hyphantria cunea, a globally invasive forest pest. In the present study, we identified and functionally characterized the Masc gene, HcMasc, in H. cunea. Sequence analysis revealed that HcMASC contained the conserved CCCH-type zinc finger domain, nuclear localization signal, and male determining domain, in which the last was confirmed to be required for its masculinization in BmN cell line. However, expression data showed that unlike male-biased expression in B. mori, HcMasc gene expresses in main all developmental stages or tissues in both sexes. Clustered regularly interspaced palindromic repeats (CRISPR) / CRISPR-associated protein 9-based disruption of the common exons 1 and 3 of the HcMasc gene resulted in imbalanced sex ratio and abnormal external genitalia of both sexes. Our results suggest that the HcMasc gene is required for both male and female sexual differentiation and dosage compensation in H. cunea and provide a foundation for developing better strategies to control this pest.
摘要:
男性化基因,Masc,编码鳞翅目特异性新型CCCH型锌指蛋白,控制家蚕的性别决定和剂量补偿。考虑到它在病虫害防治中的潜在应用,有必要研究美国白蛾Masc基因的功能,一种全球入侵森林害虫。在本研究中,我们鉴定并功能鉴定了Masc基因,HcMasc,在H.cunea。序列分析显示HcMASC含有保守的CCCH型锌指结构域,核定位信号,和男性决定领域,其中最后一个被证实是BmN细胞系中男性化所必需的。然而,表达数据显示,与男性偏向的表达不同。HcMasc基因主要表达于两性的所有发育阶段或组织中。HcMasc基因共同外显子1和3的基于规则间隔的回文重复序列(CRISPR)/CRISPR相关蛋白9的聚集性破坏导致性别比例失衡和两性外生殖器异常。我们的结果表明,HcMasc基因对于H.cunea的雄性和雌性性分化和剂量补偿都是必需的,并为制定更好的防治该害虫的策略奠定了基础。