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
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 |
| Hinterlegungsdatum: | 03 Feb 2019 20:55 |
| Letzte Änderung: | 20 Okt 2023 08:34 |
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