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A tightly regulated and adjustable CRISPR-dCas9 based AND gate in yeast

Hofmann, Anja ; Falk, Johannes ; Prangemeier, Tim ; Happel, Dominic ; Köber, Adrian ; Christmann, Andreas ; Koeppl, Heinz ; Kolmar, Harald (2019)
A tightly regulated and adjustable CRISPR-dCas9 based AND gate in yeast.
In: Nucleic Acids Research, 2018, 47 (1)
doi: 10.1093/nar/gky1191
Artikel, Zweitveröffentlichung

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Kurzbeschreibung (Abstract)

The robust and precise on and off switching of one or more genes of interest, followed by expression or repression is essential for many biological circuits as well as for industrial applications. However, many regulated systems published to date influence the viability of the host cell, show high basal expression or enable only the overexpression of the target gene without the possibility of fine regulation. Herein, we describe an AND gate designed to overcome these limitations by combining the advantages of three well established systems, namely the scaffold RNA CRISPR/dCas9 platform that is controlled by Gal10 as a natural and by LexA-ER-AD as heterologous transcription factor. We hence developed a predictable and modular, versatile expression control system. The selection of a reporter gene set up combining a gene of interest (GOI) with a fluorophore by the ribosomal skipping T2A sequence allows to adapt the system to any gene of interest without losing reporter function. In order to obtain a better understanding of the underlying principles and the functioning of our system, we backed our experimental findings with the development of a mathematical model and single-cell analysis.

Typ des Eintrags: Artikel
Erschienen: 2019
Autor(en): Hofmann, Anja ; Falk, Johannes ; Prangemeier, Tim ; Happel, Dominic ; Köber, Adrian ; Christmann, Andreas ; Koeppl, Heinz ; Kolmar, Harald
Art des Eintrags: Zweitveröffentlichung
Titel: A tightly regulated and adjustable CRISPR-dCas9 based AND gate in yeast
Sprache: Englisch
Publikationsjahr: 22 November 2019
Ort: Darmstadt
Publikationsdatum der Erstveröffentlichung: 2018
Verlag: Oxford Academic
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Nucleic Acids Research
Jahrgang/Volume einer Zeitschrift: 47
(Heft-)Nummer: 1
DOI: 10.1093/nar/gky1191
URL / URN: https://doi.org/10.1093/nar/gky1191
Herkunft: Zweitveröffentlichung aus gefördertem Golden Open Access
Kurzbeschreibung (Abstract):

The robust and precise on and off switching of one or more genes of interest, followed by expression or repression is essential for many biological circuits as well as for industrial applications. However, many regulated systems published to date influence the viability of the host cell, show high basal expression or enable only the overexpression of the target gene without the possibility of fine regulation. Herein, we describe an AND gate designed to overcome these limitations by combining the advantages of three well established systems, namely the scaffold RNA CRISPR/dCas9 platform that is controlled by Gal10 as a natural and by LexA-ER-AD as heterologous transcription factor. We hence developed a predictable and modular, versatile expression control system. The selection of a reporter gene set up combining a gene of interest (GOI) with a fluorophore by the ribosomal skipping T2A sequence allows to adapt the system to any gene of interest without losing reporter function. In order to obtain a better understanding of the underlying principles and the functioning of our system, we backed our experimental findings with the development of a mathematical model and single-cell analysis.

URN: urn:nbn:de:tuda-tuprints-84327
Sachgruppe der Dewey Dezimalklassifikatin (DDC): 500 Naturwissenschaften und Mathematik > 530 Physik
Fachbereich(e)/-gebiet(e): 18 Fachbereich Elektrotechnik und Informationstechnik
18 Fachbereich Elektrotechnik und Informationstechnik > Institut für Nachrichtentechnik > Bioinspirierte Kommunikationssysteme
18 Fachbereich Elektrotechnik und Informationstechnik > Institut für Nachrichtentechnik
05 Fachbereich Physik
05 Fachbereich Physik > Institut für Festkörperphysik (2021 umbenannt in Institut für Physik Kondensierter Materie (IPKM))
05 Fachbereich Physik > Institut für Festkörperphysik (2021 umbenannt in Institut für Physik Kondensierter Materie (IPKM)) > Statistische Physik und komplexe Systeme
07 Fachbereich Chemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie > Allgemeine Biochemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut
Hinterlegungsdatum: 03 Feb 2019 20:55
Letzte Änderung: 02 Jul 2024 23:05
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