Rebuilding evolutionary success: Using Inscripta's Onyx Digital Genome Engineering Platform to ease and enhance Adaptive Laboratory Evolution experiments

DATE:  July 28, 2020

TIME:   10:00am PDT, 1:00pm EDT

Adaptive laboratory evolution (ALE) of microbial strains is a powerful approach in biotechnology to generate chemical tolerance and gain novel biochemical insights. However, these experiments are often lengthy in their execution (weeks to decades), require strain rebuilds for validation, and can suffer from evolutionary traps. Massively parallel microbial strain engineering using Inscripta’s Onyx™ benchtop platform enabled high throughput design and build of a barcoded library of 372 individually reconstructed, previously reported ALE strain variants, 492 gene deletions, and 84 mobile element-like insertions. This library was tested against inhibitory concentrations of NaCl, coumaric acid, and isobutyric acid for rapid selection of beneficial mutations. The application of this technology to ALE variant reconstruction has allowed for an unprecedented opportunity to query a diverse population of mutations for their role in growth responses to gain new biochemical insights. The high-throughput capabilities of this strain construction platform have enabled a new paradigm in ALE experiments and offer the opportunity to do the same across countless additional applications in both basic and applied biological sciences.

Learning Objectives:

• Introduction to Inscripta’s Onyx™ Digital Genome Engineering Platform

• Example application of genome-wide strain engineering in E. coli

• Reported adaptive laboratory evolution (ALE) variants were built in high throughput and barcode tracked to enable genotyping tolerance mutations

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