Transcriptional Profiles of Diploid Mutant Apis mellifera Embryos after Knockout of csd by CRISPR/Cas9.
Transcriptional Profiles of Diploid Mutant Apis mellifera Embryos after Knockout of csd by CRISPR/Cas9.
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CRISPR/Cas9 敲除 csd 后二倍体突变体意大利蜜蜂胚胎的转录谱
DOI：
10.3390/insects12080704
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发表时间：
2021-08-06
期刊：
影响因子：
3
通讯作者：
Su S
中科院分区：
文献类型：
--
作者：
Wang X;Lin Y;Liang L;Geng H;Zhang M;Nie H;Su S
在蜜蜂中，雄蜂是单倍体，而雌蜂是二倍体，这导致了两性之间遗传物质的根本差异。为了更好地控制雄性和雌性之间的基因表达比较，通过敲除受精胚胎中的性别决定基因互补性别决定基因(CSD)，产生了二倍体突变雄性。二倍体突变体雄蜂具有雄蜂的外部形态特征和雄蜂的性腺特征。对二倍体突变胚胎和1日龄幼虫进行了RNA测序。转录组分析表明，在二倍体突变雄虫中，一些雌性偏向基因，如工蜂富集型触角(Wat)、卵黄蛋白原(Vg)和一些毒液相关基因，表达下调。相比之下，一些偏向男性的基因，如外卖和载脂蛋白III类似蛋白(A4)，上调了。此外，共表达的基因网络表明，CSD可能与无果(Fru)相互作用非常密切，女性化(Fem)可能与六角蛋白70c(He70c)有联系，而转换器-2(Tra2)可能与肌钙蛋白T(TpnT)起作用。本研究为性别分化引起的基因表达差异提供了基础性信息。相信这项研究将为进一步研究蜜蜂雄蜂和雌蜂的不同机制奠定基础。摘要在蜜蜂中，互补性别决定因子(CSD)是性别决定的主要信号。其等位基因组成在女性为杂合子，在男性为半合子或纯合子。为了探讨性别分化后雄性和雌性之间的转录组差异，排除遗传差异，用CRISPR/Cas9敲除受精胚胎中的CSD。对产蛋后24 h、48 h、72 h和96 h的二倍体突变雄蜂和来自同一受精女王的模拟处理雌蜂进行了RNA-Seq分析。在二倍体突变体的靶序列中检测到突变。二倍体突变体雄蜂具有典型的雄性形态特征和性腺特征。转录组分析表明，在二倍体突变雄虫中，一些雌性偏向基因，如工蜂富集型触角(Wat)、卵黄蛋白原(Vg)和一些毒液相关基因，表达下调。相反，一些偏向男性的基因，如外卖和载脂蛋白-III类蛋白(A4)，在二倍体突变的雄性中有更高的表达。加权基因共表达网络分析(WGCNA)表明，CSD可能与无果(Fru)、雌性化(Fem)和六角蛋白70c(He70c)、转换蛋白-2(Tra2)和肌钙蛋白T(TpnT)之间存在相互作用。本研究为进一步研究蜜蜂等膜翅目昆虫的性别二型性和发育提供了依据。
Simple Summary In honey bees, males are haploid while females are diploid, leading to a fundamental difference in genetic materials between the sexes. In order to better control the comparison of gene expression between males and females, diploid mutant males were generated by knocking out the sex-determining gene, complementary sex determiner (csd), in fertilized embryos. The diploid mutant drones had male external morphological features, as well as male gonads. RNA sequencing was performed on the diploid mutant embryos and one-day-old larvae. The transcriptome analysis showed that several female-biased genes, such as worker-enriched antennal (Wat), vitellogenin (Vg), and some venom-related genes, were down-regulated in the diploid mutant males. In contrast, some male-biased genes, like takeout and apolipophorin-III-like protein (A4), were up-regulated. Moreover, the co-expression gene networks suggested that csd might interact very closely with fruitless (fru), feminizer (fem) might have connections with hexamerin 70c (hex70c), and transformer-2 (tra2) might play roles with troponin T (TpnT). Foundational information about the differences in the gene expression caused by sex differentiation was provided in this study. It is believed that this study will pave the ground for further research on the different mechanisms between males and females in honey bees. Abstract In honey bees, complementary sex determiner (csd) is the primary signal of sex determination. Its allelic composition is heterozygous in females, and hemizygous or homozygous in males. To explore the transcriptome differences after sex differentiation between males and females, with genetic differences excluded, csd in fertilized embryos was knocked out by CRISPR/Cas9. The diploid mutant males at 24 h, 48 h, 72 h, and 96 h after egg laying (AEL) and the mock-treated females derived from the same fertilized queen were investigated through RNA-seq. Mutations were detected in the target sequence in diploid mutants. The diploid mutant drones had typical male morphological characteristics and gonads. Transcriptome analysis showed that several female-biased genes, such as worker-enriched antennal (Wat), vitellogenin (Vg), and some venom-related genes, were down-regulated in the diploid mutant males. In contrast, some male-biased genes, such as takeout and apolipophorin-III-like protein (A4), had higher expressions in the diploid mutant males. Weighted gene co-expression network analysis (WGCNA) indicated that there might be interactions between csd and fruitless (fru), feminizer (fem) and hexamerin 70c (hex70c), transformer-2 (tra2) and troponin T (TpnT). The information provided by this study will benefit further research on the sex dimorphism and development of honey bees and other insects in Hymenoptera.
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影响因子：
10.5
作者：
Dauwalder, B;Tsujimoto, S;Mattox, W
通讯作者：
Mattox, W
影响因子：
3
作者：
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影响因子：
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影响因子：
3.9
作者：
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通讯作者：
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影响因子：
3.5
作者：
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