Controlling promoter strength and regulation in Saccharomyces cerevisiae using synthetic hybrid promoters

Biotechnology and Bioengineering
John BlazeckHal Alper

Abstract

A dynamic range of well-controlled constitutive and tunable promoters are essential for metabolic engineering and synthetic biology applications in all host organisms. Here, we apply a synthetic hybrid promoter approach for the creation of strong promoter libraries in the model yeast, Saccharomyces cerevisiae. Synthetic hybrid promoters are composed of two modular components-the enhancer element, consisting of tandem repeats or combinations of upstream activation sequences (UAS), and the core promoter element. We demonstrate the utility of this approach with three main case studies. First, we establish a dynamic range of constitutive promoters and in doing so expand transcriptional capacity of the strongest constitutive yeast promoter, P(GPD) , by 2.5-fold in terms of mRNA levels. Second, we demonstrate the capacity to impart synthetic regulation through a hybrid promoter approach by adding galactose activation and removing glucose repression. Third, we establish a collection of galactose-inducible hybrid promoters that span a nearly 50-fold dynamic range of galactose-induced expression levels and increase the transcriptional capacity of the Gal1 promoter by 15%. These results demonstrate that promoters in S. cerevisiae, and po...Continue Reading

References

Oct 1, 1971·Journal of Bacteriology·J Bassel, R Mortimer
Dec 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·L Guarente, E Hoar
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·L Guarente, M Ptashne
Jan 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·H A de BoerM Vasser
Dec 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·L GuarenteP Gifford
Sep 17, 1999·Bioinformatics·J Zhu, M Q Zhang
Jul 31, 2003·Nucleic Acids Research·Ceri Van Slyke, Elizabeth J Grayhack
Aug 27, 2005·Proceedings of the National Academy of Sciences of the United States of America·Hal AlperGregory Stephanopoulos
Aug 4, 2006·Applied and Environmental Microbiology·Elke NevoigtGregory Stephanopoulos
Sep 12, 2006·Biotechnology and Bioengineering·Elke NevoigtGregory Stephanopoulos
Jun 5, 2007·Applied and Environmental Microbiology·H Wouter WisselinkAntonius J A van Maris
Jul 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Kevin F MurphyJames J Collins
Aug 19, 2007·Applied and Environmental Microbiology·Chenfeng Lu, Thomas Jeffries
Dec 6, 2008·Microbial Cell Factories·Eric J SteenJay D Keasling
Feb 13, 2010·Journal of Biomedicine & Biotechnology·Eric Young, Hal Alper
Mar 3, 2011·Molecular Systems Biology·Andrew H Babiskin, Christina D Smolke
Jul 9, 2011·Nucleic Acids Research·Andrew H Babiskin, Christina D Smolke
Sep 20, 2011·Applied and Environmental Microbiology·John BlazeckHal Alper
Dec 2, 2011·FEMS Yeast Research·Nancy A Da Silva, Sneha Srikrishnan

❮ Previous
Next ❯

Citations

Oct 12, 2012·Applied Microbiology and Biotechnology·John BlazeckHal S Alper
Dec 25, 2012·Nucleic Acids Research·Jing LiangHuimin Zhao
Jan 10, 2014·Microbial Cell Factories·Katrin WeinhandlAndrea Camattari
Apr 13, 2013·Biotechnology Advances·Sang Woo SeoGyoo Yeol Jung
Jul 6, 2014·Applied Microbiology and Biotechnology·John BlazeckHal S Alper
Feb 26, 2016·Frontiers in Microbiology·Chun-Hsien HungYuki Nakamura
Jan 7, 2015·Current Opinion in Biotechnology·John M Leavitt, Hal S Alper
Dec 25, 2015·Fungal Genetics and Biology : FG & B·James M Wagner, Hal S Alper
Dec 3, 2014·Current Opinion in Biotechnology·J Andrew JonesMattheos A G Koffas
Dec 5, 2015·ACS Synthetic Biology·Murtaza Shabbir HussainMark Blenner
Dec 20, 2015·Trends in Biotechnology·Mark J Burk, Stephen Van Dien
Sep 20, 2015·Metabolic Engineering·John BlazeckHal S Alper
Mar 7, 2013·Molecular Microbiology·Kentaro Furukawa, Stefan Hohmann
Mar 13, 2014·Biotechnology and Bioengineering·Adam J BrownDavid C James
Mar 29, 2014·Biotechnology Journal·Irina Borodina, Jens Nielsen
Dec 3, 2014·Biotechnology Advances·Tong SiHuimin Zhao
Jul 18, 2015·Nature Communications·Heidi Redden, Hal S Alper
Apr 23, 2015·Plant Science : an International Journal of Experimental Plant Biology·Meng LiJun Liu
Mar 6, 2015·Molecular Biology of the Cell·Joydeep RoyAtin K Mandal
Jan 15, 2014·Metabolic Engineering·John W K OliverShota Atsumi
Apr 15, 2015·ACS Synthetic Biology·Michael E LeeJohn E Dueber
Apr 1, 2014·Biotechnology Advances·Tsanko S GechevBernd Mueller-Roeber
Jul 23, 2014·FEMS Yeast Research·Heidi ReddenHal S Alper
Sep 10, 2014·FEMS Yeast Research·Ching-Sung TsaiYong-Su Jin
Jul 22, 2014·FEMS Yeast Research·Michael K Jensen, Jay D Keasling
May 26, 2016·Nature Communications·Yongjin J ZhouJens Nielsen
Jun 22, 2016·Nucleic Acids Research·Arun S RajkumarJay D Keasling
Jun 18, 2016·Biotechnology Journal·Amita GuptaKristala L J Prather
Jul 9, 2016·ACS Synthetic Biology·Duo LiuYing-Jin Yuan
Jul 16, 2016·ACS Synthetic Biology·Jie ZhangJay D Keasling
Oct 6, 2016·Biotechnology Letters·Irem Avcilar-Kucukgoze, Zoya Ignatova
Oct 16, 2016·Critical Reviews in Biotechnology·Xiuxia LiuZhonghu Bai
Oct 18, 2016·Nature Communications·Nathan CrookHal S Alper
Dec 25, 2016·Molecular Biotechnology·Ronald E Hector, Jeffrey A Mertens
Jan 22, 2017·ELife·Johanna LudwigsenFelix Mueller-Planitz
Jun 13, 2015·Scientific Reports·J Andrew JonesMattheos A G Koffas

❮ Previous
Next ❯

Related Concepts

Related Feeds

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.