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Custom-designed, mass silk production in genetically engineered silkworms.

Ye Yu1,2, Kai Chen1,2, Jingxia Wang3,4

  • 1Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.

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Genetically engineered silkworms produced significantly tougher silk fibers by incorporating spider and bagworm silk proteins. This breakthrough enhances silk properties for custom applications using silkworms as bioreactors.

Keywords:
Bombyx moriEumeta variegateTALENgenome editingspider silk

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Area of Science:

  • Biotechnology
  • Materials Science
  • Genetics

Background:

  • Genetically engineered silkworms are crucial for producing silk with enhanced properties, particularly through the integration of spider silk genes.
  • Existing transformation strategies face challenges with large DNA fragment integration and efficiency.

Purpose of the Study:

  • To develop and evaluate novel transformation strategies for genetically engineered silkworms to produce superior silk proteins.
  • To assess the impact of exogenous silk proteins on the mechanical and structural properties of engineered silk fibers.

Main Methods:

  • Three distinct transformation strategies were employed: TALEN-mediated fibroin light chain fusion (BmFibL-F), TALEN-mediated fibroin heavy chain replacement (BmFibH-R), and transposon-mediated transformation (BmFibH-T).
  • Exogenous silk proteins, including major ampullate spidroin 2 (MaSp2) and Eumeta variegate fibroin heavy chain (EvFibH), were introduced into silkworms.
  • Silk fiber properties were analyzed for toughness, crystallinity, and gene expression changes via RNA-seq.

Main Results:

  • The BmFibH-R strategy achieved high yields of exogenous silk proteins (51.02% MaSp2, 64.13% EvFibH) in cocoon shells.
  • Engineered silk fibers exhibited significantly enhanced toughness (∼86% in BmFibH-T, ∼80% in BmFibH-R) and increased crystallinity (∼40%).
  • The BmFibH-R system demonstrated minimal impact on endogenous gene expression.

Conclusions:

  • Novel genetic engineering strategies, particularly BmFibH-R, enable efficient production of high-performance silk proteins in silkworms.
  • The incorporation of exogenous silk proteins substantially improves silk fiber toughness and crystallinity.
  • Genetically engineered silkworms serve as effective bioreactors for custom silk production, offering a promising avenue for advanced biomaterials.