Genetically Engineered Mycelium for Biomaterials

DevelopmentBy Arun Chakravorty and Eric Holmes

• International Genetically Engineered Machines

• Standardization of genetic parts using “BioBricks”

• Development and expansion of genetic parts registry

• Annual competitions in October and November

• Developed collaboration with Ecovative.

• Company that creates Styrofoam substitute using fungal mycelium

• Hope to use synthetic biology to improve marketability

Chassis

• Ganoderma lucidum

• Higher order basidiomycete

• Sequenced in 2012• Reported to have

medicinal properties

Goals• Improve product via introduction of

several biological plasmid constructs.• Antifungal constructs to eliminate fungal contaminants

o Provide fungal species opportunity to outcompete fungal contaminants

• Carotenoid pigment pathways to make product more appealing

• Develop fungal toolkit

Homologous Recombination• Integrate linearized plasmids via natural

homologous recombination

Figure from Gene Bridges

Gibson Assembly• Used for cloning in genes with internal cut sites• Uses polymerase, 5’ exonuclease, and ligase in simultaneous reaction

Photo from Integrated DNA Technologies

Constructs

Site-Directed Mutagenesis• Introduce silent mutations• Used on genes with internal restriction sites

Image from Agilent Technologies

Thermal Cycling

DpnI Digestion

Transformation

Promoter Problems• Fungal promoters are very long

o E.coli has a check system where it splices out potential duplications

• Very few identified Ganoderma promoters• Potential solution: use T7 bacteriophage

promoter/polymerase

Image credit: Thomas Splettstoesser

Constructs

Acknowledgments

• Weill Institute• Cornell Institute for

Biotechnology & Life Sciences Technology

• College of Engineering• Turgeon Lab• Corning• Geneious• The entire Cornell iGEM

team

Thank you for listening!