Engineering transgenic silkworms to produce spider silk fibers

工程转基因蚕生产蜘蛛丝纤维

• 批准号: 7364972
• 负责人: Donald L. Jarvis
• 金额: $ 21.52万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7364972
• 关键词: Adopted; Area; Bandage; Biochemical Genetics; Bombyx; Burn injury; Condition; Cosmetic surgery; Elasticity; Engineering; Ensure; Fiber; Fibroins; Future; Genes; Genetic Transformation; Gland; Human Resources; Individual; Insecta; Knowledge; Ligaments; Light; Measures; Mechanics; Moths; Nature; Neurologic; Organ; Patients; Peptide Leader Sequences; Peptides; Production; Property; Proteins; Purpose; Recombinant Proteins; Recombinants; Recording of previous events; Research; Research Personnel; Silk; Skin graft; Spiders; Staging; Surgical sutures; Synthetic Genes; System; Tendon structure; Tensile Strength; Time; Transgenic Animals; Transgenic Organisms; base; design; experience; member; mutant; physical property; programs; promoter; protein structure function; scaffold; success; tissue regeneration; vector

DESCRIPTION (provided by applicant): Silk fibers have many current and future biomedical applications. They are widely used as fine suture materials and even thinner fibers are needed for ocular, neurological, and cosmetic surgeries. Silk fibers also hold promise as materials for artificial ligaments and tendons and they have many other potential biomedical applications. Many different recombinant protein production systems have been used to try to meet current needs and to develop additional biomedical applications of silks. Each has yielded silk proteins, but none has consistently yielded useful silk fibers. Thus, the overall purpose of this R21 exploratory proposal is to develop a system that can produce spider silk fibers. Our basic approach will be to adopt the silkworm as a surrogate host for spider silk protein production. The highly efficient piggybac system will be used to genetically transform mutant silkworms, which produce no native silk, with a synthetic gene encoding an unusually large spider silk protein. The genetic and biochemical properties of the resulting transgenic silkworms, particularly their ability to produce spider silk fibers, will then be critically assessed. A key feature of our plan is that it includes specific measures that will optimize our ability to obtain fibers. These include: (i) using a synthetic gene encoding an unusually large spider silk protein, (ii) using a promoter that targets expression of the heterologous silk proteins to the silk gland, which is naturally equipped to spin silk fibers, and (iii) using an appropriate leader peptide for secretion from the silk gland, and (iv) using a surrogate host, the silk moth, which is highly amenable to genetic transformation and naturally equipped to spin silk fibers. The successful isolation of a transgenic silkmoth that can produce spider silk fibers in this exploratory project will set the stage for future projects designed to further develop this system to produce spider silk fibers with pre-determined physical properties optimized for specific biomedical applications. This will exploit current knowledge of specific peptide motifs contributing tensile strength or elasticity to spider silks. Theoretically, these motifs can be combined in various ways to design silk fibers differing in strength and elasticity. The overall likelihood that the current project can be successfully completed is enhanced by the fact that it will be undertaken by a team of three researchers with established, complementary programs in three areas key to the project: spider silks (Lewis), insect expression systems (Jarvis), and insect transformation (Fraser). Silk fibers have many current and future biomedical applications. They are widely used as fine suture materials and even thinner fibers are needed for ocular, neurological, and cosmetic surgeries. Silk fibers also hold promise as materials for artificial ligaments and tendons and they have many other potential biomedical applications. Many different recombinant protein production systems have been used to try to meet current needs and to develop additional biomedical applications of silks. Each has yielded silk proteins, but none has consistently yielded useful silk fibers. Thus, the overall purpose of this R21 exploratory proposal is to develop a system that can produce spider silk fibers for these current and future biomedical applications. Our plan for this exploratory project is to isolate a transgenic silkworm that can produce spider silk fibers. If this project is successful, it will set the stage for future projects designed to further develop this system to produce spider silk fibers with pre-determined physical properties optimized for specific biomedical applications. These projects will exploit current knowledge of specific peptide motifs contributing tensile strength or elasticity to spider silks.

描述（由申请人提供）：丝绸纤维具有许多当前和未来的生物医学应用。它们被广泛用作精细的缝合材料，甚至需要更薄的眼部，神经系统和美容手术。丝绸纤维还具有人造韧带和肌腱的材料，并且还有许多其他潜在的生物医学应用。许多不同的重组蛋白质生产系统已被用来满足当前需求并开发丝绸的其他生物医学应用。每种都产生了丝绸蛋白质，但没有一个始终产生有用的丝纤维。因此，该R21探索性建议的总体目的是开发一种可以产生蜘蛛丝纤维的系统。我们的基本方法是将蚕作为蜘蛛丝蛋白生产的替代宿主。高效的Piggybac系统将用于转化基因蚕，而突变的蚕，没有产生天然丝，具有编码异常大的蜘蛛丝蛋白的合成基因。然后将对所得的转基因蚕的遗传和生化特性，尤其是它们产生蜘蛛丝纤维的能力，然后进行严格评估。我们计划的一个关键特征是它包括特定的措施，以优化我们获得纤维的能力。 These include: (i) using a synthetic gene encoding an unusually large spider silk protein, (ii) using a promoter that targets expression of the heterologous silk proteins to the silk gland, which is naturally equipped to spin silk fibers, and (iii) using an appropriate leader peptide for secretion from the silk gland, and (iv) using a surrogate host, the silk moth, which is highly amenable to genetic transformation and自然配备了纺丝纤维。在这个探索性项目中成功隔离可以产生蜘蛛丝纤维的转基因丝绸石，将为未来的项目奠定舞台，旨在进一步开发该系统，以生产具有针对特定生物医学应用优化的预定物理特性的蜘蛛丝纤维。这将利用当前对特定肽基序的知识，从而促进蜘蛛丝的拉伸强度或弹性。从理论上讲，这些基序可以通过各种方式组合来设计丝纤维的强度和弹性不同。当前项目可以成功完成的总体可能性通过以下事实增强，即由三个研究人员组成的团队在该项目的三个领域中建立了互补的计划：蜘蛛丝（刘易斯），昆虫表达系统（jarvis）和昆虫转化（弗雷泽（Fraser））。丝绸纤维具有许多当前和未来的生物医学应用。它们被广泛用作精细的缝合材料，甚至需要更薄的眼部，神经系统和美容手术。丝绸纤维还具有人造韧带和肌腱的材料，并且还有许多其他潜在的生物医学应用。许多不同的重组蛋白质生产系统已被用来满足当前需求并开发丝绸的其他生物医学应用。每种都产生了丝绸蛋白质，但没有一个始终产生有用的丝纤维。因此，该R21探索性建议的总体目的是开发一种可以为这些当前和将来的生物医学应用产生蜘蛛丝纤维的系统。我们对这个探索性项目的计划是隔离可以产生蜘蛛丝纤维的转基因蚕。如果该项目成功，它将为未来的项目奠定舞台，旨在进一步开发该系统，以生产具有针对特定生物医学应用优化的预定物理特性的蜘蛛丝纤维。这些项目将利用当前对特定肽图案的知识，从而促进蜘蛛丝的拉伸强度或弹性。