Design, mutate, screen: Multiplexed creation and arrayed screening of synchronized genetic clocks.

Area of Science:

• Synthetic Biology
• Genetic Engineering
• Microfluidics

Background:

• Coordinating cellular behavior via cell-cell interactions is a key goal in synthetic biology.
• Designing and screening complex genetic circuits for population-level functions is challenging.
• Existing screening methods are not compatible with population-level phenotypes dependent on cell communication.

Purpose of the Study:

• To develop an arrayed-screening workflow for dynamic, population-level genetic circuits.
• To overcome limitations of current screening technologies for cell-cell communication-dependent phenotypes.
• To enable the design and construction of novel synthetic gene circuits.

Main Methods:

• Utilized directed mutagenesis to create a mutant library of a synchronized lysis circuit.
• Employed multiplexed microfluidics for an arrayed-screening workflow.
• Developed a screening platform compatible with population-level phenotypes.

Main Results:

• Successfully screened a mutant library of an existing oscillator circuit.
• Discovered variants with distinct period-amplitude characteristics.
• Constructed a transcriptionally regulated synchronized oscillator functioning over long timescales.

Conclusions:

• The developed arrayed-screening workflow is effective for dynamic, population-level genetic circuits.
• This method facilitates the discovery and engineering of synthetic gene circuits with desired population behaviors.
• Advances synthetic biology capabilities in designing coordinated cellular functions.