» Articles » PMID: 31467340

Fine-tuning the Expression of Target Genes Using a DDI2 Promoter Gene Switch in Budding Yeast

Overview
Journal Sci Rep
Specialty Science
Date 2019 Aug 31
PMID 31467340
Citations 5
Authors
Affiliations
Soon will be listed here.
Abstract

Tuned gene expression is crucial to the proper growth and response to the environmental changes of an organism. To enable tunable gene expression as designed is desirable in both scientific research and industrial application. Here, we introduce a novel promoter switching method based on the DDI2 promoter (P) that can fine tune the expression of target genes. We constructed a recyclable cassette (P-URA3-P) and integrated it upstream of yeast target genes to replace the native promoters by DDI2 promoter without introducing any junk sequence. We found that the presence or absence of cyanamide as an inducer could turn on or off the expression of target genes. In addition, we showed that P could act as a gene switch to linearly regulate the expression levels of target genes in vivo. We switched the original promoters of RAD18, TUP1, and CDC6 with P as a proof-of-concept.

Citing Articles

Cyanamide-inducible expression of homing nuclease I for selectable marker removal and promoter characterisation in .

McDonnell L, Evans S, Lu Z, Suchoronczak M, Leighton J, Ordeniza E Synth Syst Biotechnol. 2024; 9(4):820-827.

PMID: 39072146 PMC: 11277796. DOI: 10.1016/j.synbio.2024.06.009.


Uniform [C,N]-labeled and glycosylated IgG1 Fc expressed in Saccharomyces cerevisiae.

Davis A, Roberts E, Jonathan Amster I, Barb A J Biomol NMR. 2023; 78(1):9-18.

PMID: 37989910 PMC: 11025670. DOI: 10.1007/s10858-023-00428-1.


A Modified Yeast Two-Hybrid Platform Enables Dynamic Control of Expression Intensities to Unmask Properties of Protein-Protein Interactions.

Feuer E, Zimran G, Shpilman M, Mosquna A ACS Synth Biol. 2022; 11(8):2589-2598.

PMID: 35895499 PMC: 9442787. DOI: 10.1021/acssynbio.2c00192.


De novo biosynthesis of rubusoside and rebaudiosides in engineered yeasts.

Xu Y, Wang X, Zhang C, Zhou X, Xu X, Han L Nat Commun. 2022; 13(1):3040.

PMID: 35650215 PMC: 9160076. DOI: 10.1038/s41467-022-30826-2.


GET pathway mediates transfer of mislocalized tail-anchored proteins from mitochondria to the ER.

Matsumoto S, Ono S, Shinoda S, Kakuta C, Okada S, Ito T J Cell Biol. 2022; 221(6).

PMID: 35442388 PMC: 9026240. DOI: 10.1083/jcb.202104076.

References
1.
Longtine M, McKenzie 3rd A, DeMarini D, Shah N, Wach A, Brachat A . Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast. 1998; 14(10):953-61. DOI: 10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U. View

2.
Landgraf D, Huh D, Hallacli E, Lindquist S . Scarless Gene Tagging with One-Step Transformation and Two-Step Selection in Saccharomyces cerevisiae and Schizosaccharomyces pombe. PLoS One. 2016; 11(10):e0163950. PMC: 5063382. DOI: 10.1371/journal.pone.0163950. View

3.
Butt T, Sternberg E, Gorman J, Clark P, Hamer D, Rosenberg M . Copper metallothionein of yeast, structure of the gene, and regulation of expression. Proc Natl Acad Sci U S A. 1984; 81(11):3332-6. PMC: 345501. DOI: 10.1073/pnas.81.11.3332. View

4.
MANN R, Grunstein M . Histone H3 N-terminal mutations allow hyperactivation of the yeast GAL1 gene in vivo. EMBO J. 1992; 11(9):3297-306. PMC: 556864. DOI: 10.1002/j.1460-2075.1992.tb05408.x. View

5.
Alper H, Fischer C, Nevoigt E, Stephanopoulos G . Tuning genetic control through promoter engineering. Proc Natl Acad Sci U S A. 2005; 102(36):12678-83. PMC: 1200280. DOI: 10.1073/pnas.0504604102. View