Automated Strain Construction for Biosynthetic Pathway Screening in Yeast

Area of Science:

• Synthetic Biology
• Bioengineering
• Automation and Robotics

Background:

• The Design-Build-Test-Learn (DBTL) cycle in synthetic biology is often limited by manual strain construction.
• Integrating robotic automation into synthetic biology workflows is crucial for accelerating research and development.

Purpose of the Study:

• To present a modular, integrated protocol for automating the 'Build' step in Saccharomyces cerevisiae strain construction.
• To enable high-throughput screening for pathway discovery and optimization in biofoundries.

Main Methods:

• Programming a Hamilton Microlab VANTAGE liquid handling platform to automate yeast transformations.
• Integrating off-deck hardware using a robotic arm for enhanced workflow capabilities.
• Developing a user interface with Hamilton VENUS software for customizable parameters.

Main Results:

• Achieved a throughput of 2,000 yeast transformations per week.
• Successfully screened a gene library in an engineered yeast strain for verazine biosynthesis.
• Identified pathway bottlenecks and genes enhancing verazine production by 2.0- to 5-fold.

Conclusions:

• The developed automated pipeline significantly increases throughput for yeast strain construction.
• This resource facilitates library screening for synthetic biologists in biofoundries, aiding pathway discovery, optimization, and protein engineering.