TRANSCRIPTION FACTOR Bmsage PLAYS A CRUCIAL ROLE IN SILK GLAND GENERATION IN SILKWORM, Bombyx mori
TRANSCRIPTION FACTOR Bmsage PLAYS A CRUCIAL ROLE IN SILK GLAND GENERATION IN SILKWORM, Bombyx mori
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转录因子 Bmsage 在桑蚕丝腺生成中发挥重要作用
DOI：
10.1002/arch.21244
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发表时间：
2015
影响因子：
2.2
通讯作者：
Miao Yun-gen
中科院分区：
文献类型：
--
作者：
Xin Hu-hu;Zhang Deng-pan;Chen Rui-ting;Cai Zi-zheng;Lu Yan;Liang Shuang;Miao Yun-gen
果蝇胚胎中唾液腺分泌发生改变，鼠尾草基因功能丧失。根据透射电子显微镜，唾液的体积减少，电子密度增加，导致管扩张和收缩区域，并伴有间歇性管关闭。然而，尽管 Bmsage 的序列已存储在 SilkDB 中，但 Bmsage 在蚕（Bombyx mori）中的确切功能尚不清楚。由此推断，Bmsage是一种转录因子，调节丝素蛋白的合成，并与另一种丝腺特异性转录因子，即丝腺因子-1相互作用。在这项研究中，我们利用 Cas9/sgRNA 系统（一种基因组编辑技术）引入了 Bmsage 的种系突变，导致 Bmsage 从家蚕基因组中删除。在 15 个测试样本中，有 7 个在目标位点显示出变化。诱变效率约为46.7%，未出现明显脱靶效应。筛选出的纯合突变体中，丝腺发育不良，中、后丝腺（MSG和PSG）缺失，与野生型有显着差异。 G0马赛克蚕的后代具有插入缺失突变，导致目标位点发生2或9个bp缺失，但表现出相同的异常丝腺结构。含有不同 Bmsage 开放阅读框的突变幼虫具有相同的丝腺表型。这说明突变表型是由于 Bmsage 敲除造成的。我们得出结论，Bmsage 参与丝腺的胚胎发育。
Salivary gland secretion is altered in Drosophila embryos with loss of function of the sage gene. Saliva has a reduced volume and an increased electron density according to transmission electron microscopy, resulting in regions of tube dilation and constriction with intermittent tube closure. However, the precise functions of Bmsage in silkworm (Bombyx mori) are unknown, although its sequence had been deposited in SilkDB. From this, Bmsage is inferred to be a transcription factor that regulates the synthesis of silk fibroin and interacts with another silk gland‐specific transcription factor, namely, silk gland factor‐1. In this study, we introduced a germline mutation of Bmsage using the Cas9/sgRNA system, a genome‐editing technology, resulting in deletion of Bmsage from the genome of B. mori. Of the 15 tested samples, seven displayed alterations at the target site. The mutagenesis efficiency was about 46.7% and there were no obvious off‐target effects. In the screened homozygous mutants, silk glands developed poorly and the middle and posterior silk glands (MSG and PSG) were absent, which was significantly different from the wild type. The offspring of G0mosaic silkworms had indel mutations causing 2‐ or 9‐bp deletions at the target site, but exhibited the same abnormal silk gland structure. Mutant larvae containing different open‐reading frames of Bmsage had the same silk gland phenotype. This illustrated that the mutant phenotype was due to Bmsage knockout. We conclude that Bmsage participates in embryonic development of the silk gland.