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CRISPR-based multiplexed genome editing for improved heterologous protein engineering and expression in E. coli

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Date: Monday 14 June 2021 08:00 [PT] 11:00 [ET] 16:00 [BST]

During this webinar, discover how Inscripta experts generated an Escherichia coli saturation mutagenesis library against an integrated green fluorescent protein (GFP) using CRISPR-based technology, in a few short weeks. Several engineered variants showed distinct intensity and spectral characteristics, including several literature-validated spectral variants as well as novel variants. Also uncover how a library of all predicted E. coli promoters and ribosome binding sites (RBS) 5′ of the integrated GFP start codon was generated, capable of identifying bins of low, medium and high strength under minimal and rich media.
Learn how these high-throughput profiling experiments provide excellent insights into genetic regulation and successfully deliver a measure of native sequences. The applications presented in this webinar demonstrate the power of the Onyx platform to improve heterologous-protein engineering and expression workflows while generating new biochemical insights.

What will you learn?

  • How to develop and analyze site saturation mutagenesis libraries for heterologous-enzyme engineering to improve activity
  • How to improve expression of heterologous proteins in E. coli using genome-wide editing libraries
  • Novel promoter and ribosome binding site biochemistry for improving the understanding of genome regulation
  • How to apply fluorescence-based assays for high-throughput novel variant discovery and investigations into genomic regulatory elements

Who may this interest?

  • Pharmaceutical companies with protein expression workflow in their pipeline
  • Structural biologists
  • Synthetic biologists

Speaker

Tyson Shepherd, PhD
Senior Scientist, Applications Development
Inscripta (CO, USA)

Dr Tyson Shepherd has a background in biochemistry and synthetic biology, and has published papers on protein engineering, large DNA construct design, cell-free synthetic biology, phage biology and nucleic acid nanotechnology. Dr Shepherd works in a collaborative team within Inscripta and with external partners to design and execute projects demonstrating the many applications of the Onyx Digital Genome Engineering Platform. Before joining Inscripta, Dr Shepherd was a research associate at MIT. Dr Shepherd earned his graduate degree at the University of Iowa and completed his postdoctoral research at MIT and Uppsala University.

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