A blueprint for glow tag engineering

Lee, Jihwan; St-Pierre, François

doi:10.1038/s41570-021-00351-9

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■ Automating Multiwell Plate Screening1

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2022

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“…Because the number of ways a POD could be mutated exceeds the number of atoms in the universe, 77 even pooled library methods can sample only a small fraction of the enormous sequence space. The ability to prioritize specific mutations, or at least residues to target by saturation mutagenesis, can accelerate POD engineering and increase success.…”

Section: ■ Automating Multiwell Plate Screeningmentioning

confidence: 99%

Automating the High-Throughput Screening of Protein-Based Optical Indicators and Actuators

et al. 2022

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Over the last 25 years, protein engineers have developed an impressive collection of optical tools to interface with biological systems: indicators to eavesdrop on cellular activity and actuators to poke and prod native processes. To reach the performance level required for their downstream applications, protein-based tools are usually sculpted by iterative rounds of mutagenesis. In each round, libraries of variants are made and evaluated, and the most promising hits are then retrieved, sequenced, and further characterized. Early efforts to engineer protein-based optical tools were largely manual, suffering from low throughput, human error, and tedium. Here, we describe approaches to automating the screening of libraries generated as colonies on agar, multiwell plates, and pooled populations of single-cell variants. We also briefly discuss emerging approaches for screening, including cell-free systems and machine learning.

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Section: ■ Automating Multiwell Plate Screeningmentioning

confidence: 99%