Enhanced protein and biochemical production using CRISPRi-based growth switches

Li S, Jendresen CB, Grünberger A, Ronda C, Jensen SI, Noack S, Nielsen AT (2016)
Metabolic Engineering 38: 274-284.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
Production of proteins and biochemicals in microbial cell factories is often limited by carbon and energy spent on excess biomass formation. To address this issue, we developed several genetic growth switches based on CRISPR interference technology. We demonstrate that growth of Escherichia coli can be controlled by repressing the DNA replication machinery, by targeting dnaA and oriC, or by blocking nucleotide synthesis through pyrF or thyA. This way, total GFP-protein production could be increased by up to 2.2-fold. Single-cell dynamic tracking in microfluidic systems was used to confirm functionality of the growth switches. Decoupling of growth from production of biochemicals was demonstrated for mevalonate, a precursor for isoprenoid compounds. Mass yield of mevalonate was increased by 41%, and production was maintained for more than 45 h after activation of the pyrF-based growth switch. The developed methods represent a promising approach for increasing production yield and titer for proteins and biochemicals.
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Zeitschriftentitel
Metabolic Engineering
Band
38
Seite
274-284
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Li S, Jendresen CB, Grünberger A, et al. Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering. 2016;38:274-284.
Li, S., Jendresen, C. B., Grünberger, A., Ronda, C., Jensen, S. I., Noack, S., & Nielsen, A. T. (2016). Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering, 38, 274-284. doi:10.1016/j.ymben.2016.09.003
Li, S., Jendresen, C. B., Grünberger, A., Ronda, C., Jensen, S. I., Noack, S., and Nielsen, A. T. (2016). Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering 38, 274-284.
Li, S., et al., 2016. Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering, 38, p 274-284.
S. Li, et al., “Enhanced protein and biochemical production using CRISPRi-based growth switches”, Metabolic Engineering, vol. 38, 2016, pp. 274-284.
Li, S., Jendresen, C.B., Grünberger, A., Ronda, C., Jensen, S.I., Noack, S., Nielsen, A.T.: Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering. 38, 274-284 (2016).
Li, Songyuan, Jendresen, Christian Bille, Grünberger, Alexander, Ronda, Carlotta, Jensen, Sheila Ingemann, Noack, Stephan, and Nielsen, Alex Toftgaard. “Enhanced protein and biochemical production using CRISPRi-based growth switches”. Metabolic Engineering 38 (2016): 274-284.

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System-level perturbations of cell metabolism using CRISPR/Cas9.
Jakočiūnas T, Jensen MK, Keasling JD., Curr Opin Biotechnol 46(), 2017
PMID: 28365497

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