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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Bacterial Genome Editing Strategy for Control of Transcription and Protein Stability. / Lauritsen, Ida; Martinez, Virginia; Ronda, Carlotta; Nielsen, Alex Toftgaard; Nørholm, Morten H. H.

Synthetic Metabolic Pathways. Vol. 1671 2018. p. 27-37 (Methods in Molecular Biology).

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

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@inbook{b8f1f3ee7e2149beb5fc090ab7bbe6a3,
title = "Bacterial Genome Editing Strategy for Control of Transcription and Protein Stability",
abstract = "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.",
keywords = "CRISPR interference, CRISPR-Cas9, CRMAGE, CRiPi, Essential genes, Genome editing, N-Degron, N-End rule pathway, PROTi, Protein stability",
author = "Ida Lauritsen and Virginia Martinez and Carlotta Ronda and Nielsen, {Alex Toftgaard} and N{\o}rholm, {Morten H. H.}",
year = "2018",
doi = "10.1007/978-1-4939-7295-1_3",
language = "English",
isbn = "978-1-4939-7294-4",
volume = "1671",
pages = "27--37",
booktitle = "Synthetic Metabolic Pathways",

}

RIS

TY - CHAP

T1 - Bacterial Genome Editing Strategy for Control of Transcription and Protein Stability

AU - Lauritsen, Ida

AU - Martinez, Virginia

AU - Ronda, Carlotta

AU - Nielsen, Alex Toftgaard

AU - Nørholm, Morten H. H.

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

KW - CRISPR interference

KW - CRISPR-Cas9

KW - CRMAGE

KW - CRiPi

KW - Essential genes

KW - Genome editing

KW - N-Degron

KW - N-End rule pathway

KW - PROTi

KW - Protein stability

U2 - 10.1007/978-1-4939-7295-1_3

DO - 10.1007/978-1-4939-7295-1_3

M3 - Book chapter

SN - 978-1-4939-7294-4

VL - 1671

SP - 27

EP - 37

BT - Synthetic Metabolic Pathways

ER -