scholarly journals Cas9-mediated gene editing in Saccharomyces cerevisiae

2017 ◽  
Author(s):  
Ranjith Anand ◽  
Gonen Memisoglu ◽  
James Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Editing

scholarly journals Construction of CRISPR/CAS9 system for industrial Saccharomyces cerevisiae strain and genetic manipulation effect on 2-phenylethanol pathway

2021 ◽  
Author(s):  
Zhiwei Xu ◽  
Zhe Chen ◽  
Lucheng Lin ◽  
Kun Wang ◽  
Jie Sun ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Editing ◽  
High Efficiency ◽  
Genetic Manipulation ◽  
Market Demand ◽  
Dna Homology ◽  
Ehrlich Pathway ◽  
Key Factor ◽  
Industrial Strains ◽  
Manipulation Strategy

Abstract Background The market demand for natural 2-phenylethanol (2-PE) continues to increase. Saccharomyces cerevisiae can synthesize 2-PE through the Ehrlich pathway. There are few studies on the improvement of the diploid industrial strains of S. cerevisiae by gene editing technology. There is no report on the comparison of genetic manipulation effect among S.cerevisiae strains with different 2-PE yield background, and the study on knockout of 2-PE downstream product synthesis gene and its effect on the yield of 2-PE have not been found. Results The CRISPR/CAS9 system with high efficiency for diploid S.cerevisiae CWY132 strain for industrial production of 2-PE was constructed. When the length of the homology arm of donor DNA is increased from 60bp to 500bp, the efficiency of gene editing increased from 0–100%. Using CRISPR/CAS9 technology, the branched acetaldehyde dehydrogenase genes ALD2 and ALD3 and the terminal acetyltransferase gene ATF1 in the Ehrlich pathway of S.cerevisiae strains with different 2-PE yields were knocked out. The results showed that in the high-yielding CWY-132 strain, the 2-PE yield decreased from 3.50 g/L to 1.65 g/L when double ALD2 and ALD3 were knocked out, a decrease of 52.8%. When ATF1 was knocked out, the yield of 2-PE decreased to 0.83 g/L, a decrease of 76.2%; In the low-yielding strain PK-2C, the yield of 2-PE increased from 0.21 g/L to 1.20 g/L when ALD2 was knocked out, an increase of 471%. When ATF1 was knocked out, the yield of 2-PE increased to 0.45g/L, an increase of 114%. The results show that the same genetic manipulation strategy for strains with different 2-PE yeilds backgrounds produces significantly different or even opposite effects. In addition, we found that the insufficient supply of NADH in cells can significantly affect the production of 2-PE, and the tolerance of cells to 2-PE is also a key factor that limits the further increase of 2-PE production in high-yielding strain. Conclusions This study shows that the length of the Donor DNA homology arm is a key factor affecting the efficiency of CRISPR/CAS9 gene editing in industrial diploid S. cerevisiae strains. Our result also shows that it is not feasible to increase the 2-PE production in high-yielding strains by blocking the branch pathway in the Ehrlich pathway. Breakthrough in the 2-PE yield of the high-yielding strains requires improved strains’ tolerance to 2-PE and increase the cellular NADH level.

scholarly journals Applications of CRISPR/Cas Technology to Research the Synthetic Genomics of Yeast

2021 ◽  
Author(s):  
Huafeng Lin ◽  
Haizhen Wang ◽  
Aimin Deng ◽  
Minjing Rong ◽  
Lei Ye ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Synthetic Biology ◽  
Gene Editing ◽  
Functional Genes ◽  
Whole Genome ◽  
Yeast Saccharomyces Cerevisiae ◽  
Synthetic Genome ◽  
The World ◽  
Synthetic Genomics ◽  
Genome Projects

The whole genome projects open the prelude to the diversity and complexity of biological genome by generating immense data. For the sake of exploring the riddle of the genome, scientists around the world have dedicated themselves in annotating for these massive data. However, searching for the exact and valuable information is like looking for a needle in a haystack. Advances in gene editing technology have allowed researchers to precisely manipulate the targeted functional genes in the genome by the state-of-the-art gene-editing tools, so as to facilitate the studies involving the fields of biology, agriculture, food industry, medicine, environment and healthcare in a more convenient way. As a sort of pioneer editing devices, the CRISPR/Cas systems having various versatile homologs and variants, now are rapidly giving impetus to the development of synthetic genomics and synthetic biology. Firstly, in the chapter, we will present the classification, structural and functional diversity of CRISPR/Cas systems. Then we will emphasize the applications in synthetic genome of yeast (Saccharomyces cerevisiae) using CRISPR/Cas technology based on year order. Finally, the summary and prospection of synthetic genomics as well as synthetic biotechnology based on CRISPR/Cas systems and their further utilizations in yeast are narrated.

scholarly journals Enhanced scale and scope of genome engineering and regulation using CRISPR/Cas in Saccharomyces cerevisiae

2019 ◽  
Vol 19 (7) ◽  
Author(s):  
Matthew Deaner ◽  
Hal S Alper
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Regulation ◽  
Transcriptional Regulation ◽  
Gene Editing ◽  
Genome Engineering ◽  
Guide Rna ◽  
On Demand ◽  
Large Numbers ◽  
Genetic Modifications ◽  
Genome Scale

ABSTRACT Although only 6 years old, the CRISPR system has blossomed into a tool for rapid, on-demand genome engineering and gene regulation in Saccharomyces cerevisiae. In this minireview, we discuss fundamental CRISPR technologies, tools to improve the efficiency and capabilities of gene targeting, and cutting-edge techniques to explore gene editing and transcriptional regulation at genome scale using pooled approaches. The focus is on applications to metabolic engineering with topics including development of techniques to edit the genome in multiplex, tools to enable large numbers of genetic modifications using pooled single-guide RNA libraries and efforts to enable programmable transcriptional regulation using endonuclease-null Cas enzymes.

scholarly journals A new strategy for seamless gene editing and marker recycling in Saccharomyces cerevisiae using lethal effect of Cwp1

2018 ◽  
Vol 8 (6) ◽  
Author(s):  
Yuxiao Hu ◽  
Yanrong Jia ◽  
Xiangdong Zhao ◽  
Zihao Yang ◽  
Zhimin Hao ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Editing ◽  
Lethal Effect ◽  
Marker Recycling ◽  
New Strategy

scholarly journals Reducing phenolic off-flavors through CRISPR-based gene editing of the FDC1 gene in Saccharomyces cerevisiae x Saccharomyces eubayanus hybrid lager beer yeasts

PLoS ONE ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. e0209124 ◽  
Author(s):  
Stijn Mertens ◽  
Brigida Gallone ◽  
Jan Steensels ◽  
Beatriz Herrera-Malaver ◽  
Jeroen Cortebeek ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Editing ◽  
Off Flavors ◽  
Lager Beer ◽  
Saccharomyces Eubayanus

scholarly journals Correction: Reducing phenolic off-flavors through CRISPR-based gene editing of the FDC1 gene in Saccharomyces cerevisiae x Saccharomyces eubayanus hybrid lager beer yeasts

PLoS ONE ◽  
2019 ◽  
Vol 14 (10) ◽  
pp. e0224525
Author(s):  
Stijn Mertens ◽  
Brigida Gallone ◽  
Jan Steensels ◽  
Beatriz Herrera-Malaver ◽  
Jeroen Cortebeek ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Editing ◽  
Off Flavors ◽  
Lager Beer ◽  
Saccharomyces Eubayanus

Anti-Saccharomyces cerevisiae Antibodies in Inflammatory Bowel Disease: a Family Study

2001 ◽  
Vol 36 (2) ◽  
pp. 196-201 ◽  
Author(s):  
F. Seibold ◽  
O. Stich ◽  
R. Hufnagl ◽  
S. Kamil ◽  
M. Scheurlen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Inflammatory Bowel Disease ◽  
Bowel Disease ◽  
Family Study ◽  
Inflammatory Bowel
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