Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG

Binder D, Grünberger A, Loeschcke A, Probst C, Bier C, Pietruszka J, Wiechert W, Kohlheyer D, Jaeger K-E, Drepper T (2014)
Integrative biology 6(8): 755-765.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
OA 5.86 MB
Binder, Dennis; Grünberger, AlexanderUniBi; Loeschcke, Anita; Probst, Christopher; Bier, Claus; Pietruszka, Jörg; Wiechert, Wolfgang; Kohlheyer, Dietrich; Jaeger, Karl-Erich; Drepper, Thomas
Abstract / Bemerkung
Light can be used to control numerous cellular processes including protein function and interaction as well as gene expression in a non-invasive fashion and with unprecedented spatiotemporal resolution. However, for chemical phototriggers tight, gradual, and homogeneous light response has never been attained in living cells. Here, we report on a light-responsive bacterial T7 RNA polymerase expression system based on a photocaged derivative of the inducer molecule isopropyl-β-D-thiogalactopyranoside (IPTG). We have comparatively analyzed different Escherichia coli lac promoter-regulated expression systems in batch and microfluidic single-cell cultivation. The lacY-deficient E. coli strain Tuner(DE3) harboring additional plasmid-born copies of the lacI gene exhibited a sensitive and defined response to increasing IPTG concentrations. Photocaged IPTG served as a synthetic photo-switch to convert the E. coli system into an optogenetic expression module allowing for precise and gradual light-triggering of gene expression as demonstrated at the single cell level. Graphical abstract: Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG
Integrative biology
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Binder D, Grünberger A, Loeschcke A, et al. Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG. Integrative biology. 2014;6(8):755-765.
Binder, D., Grünberger, A., Loeschcke, A., Probst, C., Bier, C., Pietruszka, J., Wiechert, W., et al. (2014). Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG. Integrative biology, 6(8), 755-765. doi:10.1039/c4ib00027g
Binder, D., Grünberger, A., Loeschcke, A., Probst, C., Bier, C., Pietruszka, J., Wiechert, W., Kohlheyer, D., Jaeger, K. - E., and Drepper, T. (2014). Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG. Integrative biology 6, 755-765.
Binder, D., et al., 2014. Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG. Integrative biology, 6(8), p 755-765.
D. Binder, et al., “Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG”, Integrative biology, vol. 6, 2014, pp. 755-765.
Binder, D., Grünberger, A., Loeschcke, A., Probst, C., Bier, C., Pietruszka, J., Wiechert, W., Kohlheyer, D., Jaeger, K.-E., Drepper, T.: Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG. Integrative biology. 6, 755-765 (2014).
Binder, Dennis, Grünberger, Alexander, Loeschcke, Anita, Probst, Christopher, Bier, Claus, Pietruszka, Jörg, Wiechert, Wolfgang, Kohlheyer, Dietrich, Jaeger, Karl-Erich, and Drepper, Thomas. “Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG”. Integrative biology 6.8 (2014): 755-765.
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