CRISPR Disruption of BmOvo Resulted in the Failure of Emergence and Affected the Wing and Gonad Development in the Silkworm Bombyx mori
CRISPR Disruption of BmOvo Resulted in the Failure of Emergence and Affected the Wing and Gonad Development in the Silkworm Bombyx mori
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CRISPR 破坏 BmOvo 导致家蚕羽化失败并影响翅膀和性腺发育
DOI：
10.3390/insects10080254
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发表时间：
2019-08-01
期刊：
影响因子：
3
通讯作者：
Xu, Jun
中科院分区：
文献类型：
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作者：
Bi, Honglun;Xu, Xia;Xu, Jun
家蚕是一种重要的经济昆虫，广泛用作鳞翅目昆虫模型。虽然家蚕的体细胞性别决定已被很好地描述，但种系性别决定还没有被描述。在这里，我们使用基于转基因的CRISPR/Cas9基因组编辑系统来研究家蚕Ovo基因的功能。BmOvo是果蝇生殖系性别决定中一个重要因子的同源物。BmOvo突变体具有异常形状的卵，其在卵巢管中紊乱，并且性腺发育异常。有趣的是，翅盘和翅膀没有正常发育，大多数突变体未能羽化。qRT-PCR分析表明，BmOvo基因在翅盘和表皮中表达量较高。与野生型动物相比，BmOvo突变体中参与WNT信号通路的基因和翅膀发育基因BmWCP 10和BmE 74下调。这些结果表明，BmOvo基因产物在翅变态中起着重要作用。因此，这项研究为BmOvo在生殖系性别决定之外的多种功能提供了新的见解。
The domesticated silkworm is an economically important insect that is widely used as a lepidopteran insect model. Although somatic sex determination in the silkworm is well characterized, germline sex determination is not. Here, we used the transgenic-based CRISPR/Cas9 genome editing system to study the function of the Ovo gene in Bombyx mori. BmOvo is the homolog of a factor important in germline sex determination in Drosophila melanogaster. BmOvo mutants had abnormally shaped eggs that were disordered in the ovarioles, and gonad development was abnormal. Interestingly, wing discs and wings did not develop properly, and most of the mutants failed to eclose. Gene expression analyses by qRT-PCR showed that BmOvo gene was highly expressed in the wing disc and epidermis. Genes involved in the WNT signaling pathway and wing development genes BmWCP10 and BmE74 were downregulated in the BmOvo mutants when compared with wild-type animals. These results demonstrate that the BmOvo gene product plays an important role in wing metamorphosis. Thus, this study provides new insights into the multiple functions of BmOvo beyond germline sex determination.