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I Grew Real Spider Silk Using Yeast

The Thought Emporium・33 minutes read

The project successfully designed yeast to produce spider silk with enhanced properties using a synthetic biology approach, overcoming initial challenges with DNA isolation from spiders. Through a meticulous process involving gene synthesis, modification, and insertion into yeast, successful production and secretion of silk were achieved, paving the way for scaling up production and further experimentation with bio-mineralization peptides and colorful proteins.

Insights

  • Designing spider silk DNA from scratch was a pivotal strategy after failed attempts to isolate DNA from spiders, highlighting the project's innovative approach to creating a new version of the silk project.
  • The integration of biomineralization peptides into the silk gene, alongside community feedback-driven modifications to the yeast protocol, underscored a meticulous process that optimized silk properties and production efficiency, showcasing a commitment to enhancing silk through advanced genetic engineering techniques.

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Recent questions

  • How is spider silk produced?

    Spider silk is produced using yeast.

  • What are the properties of spider silk?

    Spider silk is known for its strength and elasticity.

  • How is silk DNA synthesized?

    Silk DNA is synthesized by combining subunits and creating varied amino acid sequences.

  • What is the role of yeast in silk production?

    Yeast is used to produce silk proteins.

  • What are the future plans for silk production?

    Future plans involve scaling up production, testing on silk cocoons, and utilizing a bioreactor for continuous silk production.

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Summary

00:00

"Yeast tube produces spider silk DNA"

  • The yeast tube containing spider silk was a result of extensive design time, lab work, and financial investment.
  • The project aimed to use yeast to produce spider silk, specifically from the southern black widow spider.
  • Initial attempts to isolate DNA from spiders failed due to ineffective preservation methods.
  • The project pivoted to designing DNA from scratch, with a focus on creating a new version of the silk project.
  • Spider silk consists of various types, with dragline silk being the most sought-after for its strength and elasticity.
  • Synthetic biologists typically replicate silk by copying and pasting monomers to create a protein sequence.
  • The process of synthesizing silk DNA is challenging due to repeating sequences, requiring manual effort.
  • To introduce randomness while maintaining protein structure, subunits from different silk types were combined.
  • A chart of codons was created to ensure compatibility with the yeast species used for the project.
  • Random number generators were employed to create varied amino acid sequences for each subunit, ensuring diversity in the final DNA sequence.

13:46

Designing Silk Gene for Yeast Production Success

  • The designer experimented with various spacings until finding an appealing design, similar to the process of designing the SR-71 aircraft.
  • The designer aimed to create a silk-like material with specific properties, willing to adjust the design if needed.
  • Gene Universal was chosen for gene synthesis due to their variety of DNA backbones suitable for yeast, like the PPIC 9k backbone.
  • The PPIC 9k backbone was selected for its compatibility with yeast, use of G418 for selection, integration into yeast genome, and a tag for silk excretion.
  • Biomineralization peptides were added to the silk gene to enhance properties like mineralization, antimicrobial effects, and strength.
  • The silk gene with biomineralization peptides was sent for synthesis to enhance silk properties.
  • Errors in peptide sequences were discovered post-design, but fixable due to added restriction sites for easy modification.
  • The synthesized DNA was received after four months, requiring insertion into Pichia pastoris yeast, known for protein production.
  • Modifications to the yeast protocol were made based on community feedback, improving efficiency and success rates.
  • Tests confirmed successful modification of yeast to produce silk, with visual and PCR evidence supporting the presence of the silk gene and protein production.

27:38

Graphene-binding peptide enhances silk production efficiency.

  • The addition of a graphene binding peptide to the silk gene allows for easy dispersion of the silk solution, causing clumping and coagulation when mixed with graphene, while plain yeast leaves the graphene in solution.
  • After mixing the yeast samples with their respective graphene solutions, the silk solution quickly formed tiny clumps, coagulated, and sank in the tube, leaving a clear graphene-free solution above, indicating successful silk production and secretion function.
  • Future plans involve scaling up the production by growing a large batch of yeast, testing the process on silk cocoons, and using a bioreactor for continuous silk production, with the goal of making fibers and testing bio-mineralization peptides.
  • Further experiments include using a wet spinning apparatus, modifying the plasmid to produce colorful proteins for different colored silk fibers, and setting up an online store to sell DNA designs, with a focus on computational biology courses from Brilliant for learning protein design intricacies.
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