The Masculinizer gene,Masc,encodes a lepidopteran-specific novel CCCHtype zinc finger protein,which controls sex determination and dosage compensation in Bombyx mori.Considering the potential application of it in pest...The Masculinizer gene,Masc,encodes a lepidopteran-specific novel CCCHtype 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,Hc-Masc 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.展开更多
Sex determination has been studied in the model lepidopteran species Bombyx mori, but it remains poorly understood in lepidopteran pests. In the present study, we identified and characterized the Masculinizer (Masc) g...Sex determination has been studied in the model lepidopteran species Bombyx mori, but it remains poorly understood in lepidopteran pests. In the present study, we identified and characterized the Masculinizer (Masc) gene in a Noctuidae pest species, Agrotis ipsilon. Sequence analysis revealed that AiMasc encodes a protein of 658 amino acids that has two CCCH-type zinc finger domains and two conserved cysteine residues (Cys-277 and Cys-280). We assessed the masculinizing activity AiMasc in BmN cells and found that/z'Masc induced expression of the male-specific doublesex isoform. Disruption of Masc via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) in A. ipsilon caused abnormalities in abdominal segments and external genitalia, resulting in male-specific sterility. These results suggest that Masc participates in the process of sex determination in A. ipsilon. Successful identification of sex-determination gene in a pest species may enable the development of novel genetic approaches for pest control.展开更多
昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆...昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆虫性别决定是通过级联反应调控的,在鳞翅目昆虫ZW/ZZ系统中,位于W染色体上的性别决定初始信号沉默雄性化关键基因Masculinizer(Masc),然后Doublesex(dsx)基因被雌性特异性剪接,从而调控雌性偏向基因的表达;而雄虫中Masc正常表达,调控dsx进行雄性特异剪接,实现雄性分化。本文对鳞翅目昆虫性别决定初始信号、Masc和dsx的研究现状,以及基于基因编辑技术和性别决定机制在鳞翅目昆虫遗传调控中的应用进行了综述。相关研究为鳞翅目昆虫的性别筛选、不育昆虫释放技术(sterile insect technique,SIT)和释放携带雌性特异显性致死基因昆虫技术(female specific release of insects carrying dominant lethal,fsRIDL)奠定理论基础。展开更多
Secretory carcinoma（SC）, previously described as mammary analogue secretory carcinoma（MASC）, is a recently described salivary gland tumor which morphologically resembles mammary secretory carcinoma. The first desc...Secretory carcinoma（SC）, previously described as mammary analogue secretory carcinoma（MASC）, is a recently described salivary gland tumor which morphologically resembles mammary secretory carcinoma. The first description of SC/MASC, reported by Skálová et al. in 2010, was as a rare salivary carcinoma imitating secretory carcinoma of the breast. SC/MASC is a unique salivary gland tumor with morphological overlap with acinic cell carcinoma（Aci CC）, mucoepidermoid carcinoma（MEC）, and adenocarcinoma not otherwise specified（ADCNOS）. SC/MASC shares similar clinicopathological features with Aci CC. As a critical difference between SC/MASC and Aci CC, SC/MASC characteristically has the chromosomal translocation t（12;15）（p13;q25） which leads to a fusion gene between the ETV6 gene on chromosome 12 and the NTRK3 gene on chromosome 15. This genetic background is an important differential diagnostic finding for excluding other salivary gland tumors and may be a critical factor determining the prognosis for patients with SC/MASC. Research in recent years has provided a large body of new data on SC/MASC and suggests the possibility that the ETV6-NTRK3 translocation could be a therapeutic target. Here, we review the morphological and clinicopathological features of SC/MASC and discuss new directions for therapy.展开更多
MicroRNAs(miRNAs)are regulatory RNA molecules that bind to target messenger RNAs(mRNAs)and affect the stability or translational efficiency of the bound mRNAs.Single or dual-luciferase reporter systems have been succe...MicroRNAs(miRNAs)are regulatory RNA molecules that bind to target messenger RNAs(mRNAs)and affect the stability or translational efficiency of the bound mRNAs.Single or dual-luciferase reporter systems have been successfully used to identify miRNA target genes in mammalian cells.These reporter systems,however,are not sensitive enough to verify miRNA-target gene relationships in insect cell lines because the promoters of the target luciferase(usually Renilla)used in these reporter systems are too weak to drive sufficient expression of the target luciferase in insect cells.In this study,we replaced the SV40 promoter in the psiCHECK-2 reporter vector,which is widely used with mammalian cell lines,with the HSV-TK or AC5.1 promoter to yield two new dual-luciferase reporter vectors,designated psiCHECK-2-TK and psiCHECK-2-AC5.1,respectively.Only psiCHECK-2 and psiCHECK-2-AC5.1 had suitable target(7?enz7/a)/reference(firefly)luciferase activity ratios in mammalian(HeLa and HEK293)and insect(Sf9,S2,Helicoverpa zea fat body and ovary)cell lines,while psiCHECK-2-TK had suitable Renilla/firefly luciferase activity ratios regardless of the cell line.Moreover,psiCHECK-2-TK successfully detected the interaction between Helicoverpa armigera miRNA9a and its target,the 3'-untranslated region of heat shock protein 90,in both mammalian and H.zea cell lines,but psiCHECK-2 failed to do so in IT.zea cell lines.Furthermore,psiCHECK-2-TK with the target sequence,HzMasc(H.zea Masculinizer),accurately differentiated between H.zea cell lines with or without the negative regulation factor(miRNA or piRNA)of HzMasc.These data demonstrate that psiCHECK-2-TK can be used to functionally characterize small RNA target genes in both mammalian and insect cells.展开更多
基金funded by the Strategic Priority Research Program of Chinese Academy of Sciences(XDB11010600)National Natural Science Foundation of China(32101531 and 31420103918)National Key Research and Development Program of China,grant number 2018YFD0600203.
文摘The Masculinizer gene,Masc,encodes a lepidopteran-specific novel CCCHtype 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,Hc-Masc 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.
基金the National Basic Research Program of China (2015CB755703)the National Science Foundation of China (31522053,31501648,3142010391 & and 91631103)the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB11010600).
文摘Sex determination has been studied in the model lepidopteran species Bombyx mori, but it remains poorly understood in lepidopteran pests. In the present study, we identified and characterized the Masculinizer (Masc) gene in a Noctuidae pest species, Agrotis ipsilon. Sequence analysis revealed that AiMasc encodes a protein of 658 amino acids that has two CCCH-type zinc finger domains and two conserved cysteine residues (Cys-277 and Cys-280). We assessed the masculinizing activity AiMasc in BmN cells and found that/z'Masc induced expression of the male-specific doublesex isoform. Disruption of Masc via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) in A. ipsilon caused abnormalities in abdominal segments and external genitalia, resulting in male-specific sterility. These results suggest that Masc participates in the process of sex determination in A. ipsilon. Successful identification of sex-determination gene in a pest species may enable the development of novel genetic approaches for pest control.
文摘昆虫性别决定机制复杂多样,研究昆虫性别决定机制既能加强对物种进化关系的理解,也能为通过遗传调控控制害虫种群提供新方向。鳞翅目昆虫多为雌雄二型,繁殖方式以两性生殖为主,性染色体系统大多为ZW/ZZ系统,雌性为ZW型,雄性为ZZ型。昆虫性别决定是通过级联反应调控的,在鳞翅目昆虫ZW/ZZ系统中,位于W染色体上的性别决定初始信号沉默雄性化关键基因Masculinizer(Masc),然后Doublesex(dsx)基因被雌性特异性剪接,从而调控雌性偏向基因的表达;而雄虫中Masc正常表达,调控dsx进行雄性特异剪接,实现雄性分化。本文对鳞翅目昆虫性别决定初始信号、Masc和dsx的研究现状,以及基于基因编辑技术和性别决定机制在鳞翅目昆虫遗传调控中的应用进行了综述。相关研究为鳞翅目昆虫的性别筛选、不育昆虫释放技术(sterile insect technique,SIT)和释放携带雌性特异显性致死基因昆虫技术(female specific release of insects carrying dominant lethal,fsRIDL)奠定理论基础。
文摘Secretory carcinoma（SC）, previously described as mammary analogue secretory carcinoma（MASC）, is a recently described salivary gland tumor which morphologically resembles mammary secretory carcinoma. The first description of SC/MASC, reported by Skálová et al. in 2010, was as a rare salivary carcinoma imitating secretory carcinoma of the breast. SC/MASC is a unique salivary gland tumor with morphological overlap with acinic cell carcinoma（Aci CC）, mucoepidermoid carcinoma（MEC）, and adenocarcinoma not otherwise specified（ADCNOS）. SC/MASC shares similar clinicopathological features with Aci CC. As a critical difference between SC/MASC and Aci CC, SC/MASC characteristically has the chromosomal translocation t（12;15）（p13;q25） which leads to a fusion gene between the ETV6 gene on chromosome 12 and the NTRK3 gene on chromosome 15. This genetic background is an important differential diagnostic finding for excluding other salivary gland tumors and may be a critical factor determining the prognosis for patients with SC/MASC. Research in recent years has provided a large body of new data on SC/MASC and suggests the possibility that the ETV6-NTRK3 translocation could be a therapeutic target. Here, we review the morphological and clinicopathological features of SC/MASC and discuss new directions for therapy.
基金This work was supported by the National Science Foundation of China(NSFC)-Henan Joint major grant(U2004206)the State Key Laboratory of Cotton Biology(CB2020A06).
文摘MicroRNAs(miRNAs)are regulatory RNA molecules that bind to target messenger RNAs(mRNAs)and affect the stability or translational efficiency of the bound mRNAs.Single or dual-luciferase reporter systems have been successfully used to identify miRNA target genes in mammalian cells.These reporter systems,however,are not sensitive enough to verify miRNA-target gene relationships in insect cell lines because the promoters of the target luciferase(usually Renilla)used in these reporter systems are too weak to drive sufficient expression of the target luciferase in insect cells.In this study,we replaced the SV40 promoter in the psiCHECK-2 reporter vector,which is widely used with mammalian cell lines,with the HSV-TK or AC5.1 promoter to yield two new dual-luciferase reporter vectors,designated psiCHECK-2-TK and psiCHECK-2-AC5.1,respectively.Only psiCHECK-2 and psiCHECK-2-AC5.1 had suitable target(7?enz7/a)/reference(firefly)luciferase activity ratios in mammalian(HeLa and HEK293)and insect(Sf9,S2,Helicoverpa zea fat body and ovary)cell lines,while psiCHECK-2-TK had suitable Renilla/firefly luciferase activity ratios regardless of the cell line.Moreover,psiCHECK-2-TK successfully detected the interaction between Helicoverpa armigera miRNA9a and its target,the 3'-untranslated region of heat shock protein 90,in both mammalian and H.zea cell lines,but psiCHECK-2 failed to do so in IT.zea cell lines.Furthermore,psiCHECK-2-TK with the target sequence,HzMasc(H.zea Masculinizer),accurately differentiated between H.zea cell lines with or without the negative regulation factor(miRNA or piRNA)of HzMasc.These data demonstrate that psiCHECK-2-TK can be used to functionally characterize small RNA target genes in both mammalian and insect cells.
