Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine

Kusen PM, Wandrey G, Probst C, Grünberger A, Holz M, Meyer zu Berstenhorst S, Kohlheyer D, Büchs J, Pietruszka J (2016)
ACS chemical biology 11(10): 2915-2922.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
Light-mediated gene expression enables the noninvasive regulation of cellular functions. Apart from their classical application of regulating single cells with high spatiotemporal resolution, we highlight the potential of light-mediated gene expression for biotechnological issues. Here, we demonstrate the first light-mediated gene regulation in Saccharomyces cerevisiae using the repressible pMET17 promoter and the photolabile NVOC methionine that releases methionine upon irradiation with UVA light. In this system, the expression can be repressed upon irradiation and is reactivated due to consumption of methionine. The photolytic release allows precise control over the methionine concentration and therefore over the repression duration. Using this light regulation mechanism, we were able to apply an in-house constructed 48-well cultivation system which allows parallelized and automated irradiation programs as well as online detection of fluorescence and growth. This system enables screening of multiple combinations of several repression/derepression intervals to realize complex expression programs (e.g., a stepwise increase of temporally constant expression levels, linear expression rates with variable slopes, and accurate control over the expression induction, although we used a repressible promoter.) Thus, we were able to control all general parameters of a gene expression experiment precisely, namely start, pause, and stop at desired time points, as well as the ongoing expression rate. Furthermore, we gained detailed insights into single-cell expression dynamics with spatiotemporal resolution by applying microfluidics cultivation technology combined with fluorescence time-lapse microscopy.
Erscheinungsjahr
Zeitschriftentitel
ACS chemical biology
Band
11
Zeitschriftennummer
10
Seite
2915 - 2922
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Kusen PM, Wandrey G, Probst C, et al. Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine. ACS chemical biology. 2016;11(10):2915-2922.
Kusen, P. M., Wandrey, G., Probst, C., Grünberger, A., Holz, M., Meyer zu Berstenhorst, S., Kohlheyer, D., et al. (2016). Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine. ACS chemical biology, 11(10), 2915-2922. doi:10.1021/acschembio.6b00462
Kusen, P. M., Wandrey, G., Probst, C., Grünberger, A., Holz, M., Meyer zu Berstenhorst, S., Kohlheyer, D., Büchs, J., and Pietruszka, J. (2016). Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine. ACS chemical biology 11, 2915-2922.
Kusen, P.M., et al., 2016. Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine. ACS chemical biology, 11(10), p 2915-2922.
P.M. Kusen, et al., “Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine”, ACS chemical biology, vol. 11, 2016, pp. 2915-2922.
Kusen, P.M., Wandrey, G., Probst, C., Grünberger, A., Holz, M., Meyer zu Berstenhorst, S., Kohlheyer, D., Büchs, J., Pietruszka, J.: Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine. ACS chemical biology. 11, 2915-2922 (2016).
Kusen, Peter M., Wandrey, Georg, Probst, Christopher, Grünberger, Alexander, Holz, Martina, Meyer zu Berstenhorst, Sonja, Kohlheyer, Dietrich, Büchs, Jochen, and Pietruszka, Jörg. “Optogenetic Regulation of Tunable Gene Expression in Yeast Using Photo-Labile Caged Methionine”. ACS chemical biology 11.10 (2016): 2915-2922.

2 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Light-controlled gene expression in yeast using photocaged Cu2.
Kusen PM, Wandrey G, Krewald V, Holz M, Berstenhorst SMZ, Büchs J, Pietruszka J., J Biotechnol 258(), 2017
PMID: 28455204
Production of amino acids - Genetic and metabolic engineering approaches.
Lee JH, Wendisch VF., Bioresour Technol 245(pt b), 2017
PMID: 28552565

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