Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial
Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial
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DOI：
10.1073/pnas.1703449114
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发表时间：
2017-06-27
影响因子：
11.1
通讯作者：
Kotani, Eiji
中科院分区：
文献类型：
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作者：
Otsuki, Ryosuke;Yamamoto, Masafumi;Kotani, Eiji
具有特定组织功能障碍的基因操纵生物体对于各种生物学应用和机制的研究至关重要。然而，具有组织特异性功能障碍的模式生物的生物工程尚未取得进展，因为首先需要克服在个体生物的活细胞中表达蛋白质（例如细胞毒素）的挑战。在这里，我们报告了一种具有后部丝腺（PSG）的转基因蚕（Bombyx mori）的建立，该蚕旨在表达甘蓝蝴蝶（Pieris rapae）细胞毒素pierisin-1A（P1 A）。P1 A是凋亡诱导剂pierisin-1的同源物，具有比pierisin-1相对较低的DNA ADP核糖基转移酶活性;当在B中表达时，它还诱导某些蛋白质合成的抑制。Mori衍生的培养细胞。转基因衍生的具有酶活性的P1 A结构域在转基因家蚕PSG中成功表达。腺体未显示出与骨化相关的形态学变化;然而，异常外观明显。引入的截短的P1 A导致PSG功能障碍，因为它们不能产生丝蛋白丝素蛋白。由蚕产生的茧仅由胶样糖蛋白丝胶蛋白组成，可溶性丝胶蛋白可以制备成水凝胶。胚胎干细胞可以在水凝胶上维持未分化状态，并通过引入水凝胶中的细胞因子的刺激而增殖。因此，具有靶向P1 A表达的生物工程成功地产生了具有重要应用潜力的生物有用性状的蚕。
Genetically manipulated organisms with dysfunction of specific tissues are crucial for the study of various biological applications and mechanisms. However, the bioengineering of model organisms with tissue-specific dysfunction has not progressed because the challenges of expression of proteins, such as cytotoxins, in living cells of individual organisms need to be overcome first. Here, we report the establishment of a transgenic silkworm (Bombyx mori) with posterior silk glands (PSGs) that was designed to express the cabbage butterfly (Pieris rapae) cytotoxin pierisin-1A (P1A). P1A, a homolog of the apoptosis inducer pierisin-1, had relatively lower DNA ADP ribosyltransferase activity than pierisin-1; it also induced the repression of certain protein synthesis when expressed in B. mori-derived cultured cells. The transgene-derived P1A domain harboring enzymatic activity was successfully expressed in the transgenic silkworm PSGs. The glands showed no apoptosis-related morphological changes; however, an abnormal appearance was evident. The introduced truncated P1A resulted in the dysfunction of PSGs in that they failed to produce the silk protein fibroin. Cocoons generated by the silkworms solely consisted of the glue-like glycoprotein sericin, from which soluble sericin could be prepared to form hydrogels. Embryonic stem cells could be maintained on the hydrogels in an undifferentiated state and proliferated through stimulation by the cytokines introduced into the hydrogels. Thus, bioengineering with targeted P1A expression successfully produced silkworms with a biologically useful trait that has significant application potential.