Bacterial Genome Editing Strategy for Control of Transcription and Protein Stability

Research output: Chapter in Book/Report/Conference proceedingBook chapter – Annual report year: 2018Researchpeer-review

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In molecular biology and cell factory engineering, tools that enable control of protein production and stability are highly important. Here, we describe protocols for tagging genes in Escherichia coli allowing for inducible degradation and transcriptional control of any soluble protein of interest. The underlying molecular biology is based on the two cross-kingdom tools CRISPRi and the N-end rule for protein degradation. Genome editing is performed with the CRMAGE technology and randomization of the translational initiation region minimizes the polar effects of tag insertion. The approach has previously been applied for targeting proteins originating from essential operon-located genes and has potential to serve as a universal synthetic biology tool.
Original languageEnglish
Title of host publicationSynthetic Metabolic Pathways
Publication date2018
ISBN (Print)978-1-4939-7294-4
ISBN (Electronic)978-1-4939-7295-1
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • CRISPR interference, CRISPR-Cas9, CRMAGE, CRiPi, Essential genes, Genome editing, N-Degron, N-End rule pathway, PROTi, Protein stability

ID: 140018112