scholarly journals CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art

10.3791/59350 ◽  
2019 ◽  
Author(s):  
Klaudia Ciurkot ◽  
Brenda Vonk ◽  
Thomas E. Gorochowski ◽  
Johannes A. Roubos ◽  
René Verwaal
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Editing ◽  
The Creation

The creation of an expression vector for genome editing of the EDS 1 gene

Biomics ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 422-429
Author(s):  
N.A. Rozhnova ◽  
G.A. Gerashchenkov ◽  
A.V. Chemeris
Keyword(s):  
Genome Editing ◽  
Expression Vector ◽  
The Creation

scholarly journals A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Yueping Zhang ◽  
Juan Wang ◽  
Zibai Wang ◽  
Yiming Zhang ◽  
Shuobo Shi ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Editing

scholarly journals RABBIT BIOMODELS OF HUMAN DISEASES DEVELOPED USING NEW GENOMIC TECHNOLOGIES. CRISPR/CAS9 (REVIEW)

Biomeditsina ◽  
2019 ◽  
pp. 12-33
Author(s):  
E. M. Koloskova ◽  
V. N. Karkischenko ◽  
V. A. Yezersky ◽  
N. V. Petrova ◽  
S. V. Maksimenko ◽  
...  
Keyword(s):  
Genome Editing ◽  
Human Disease ◽  
Genetically Modified ◽  
Human Diseases ◽  
Present Review ◽  
Disease Models ◽  
Genomic Technologies ◽  
Highly Effective ◽  
The Creation

With the advent of endonuclease methods of genome editing, particularly CRISPR/Cas9, it has become possible to obtain genetically modified rabbits by microinjection of zygotes. These highly effective human disease models can be used for various purposes. The present review aims to consider modern achievements in the creation of rabbit biomodels of human diseases using the technologies of genetic editing. It is concluded that Russian laboratories should intensify research in the development of genetically modified rabbits that can be used for various biomedical studies and biomodelling.

scholarly journals EngineeringKluyveromyces marxianusas a Robust Synthetic Biology Platform Host

mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Paul Cernak ◽  
Raissa Estrela ◽  
Snigdha Poddar ◽  
Jeffrey M. Skerker ◽  
Ya-Fang Cheng ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Synthetic Biology ◽  
Genome Editing ◽  
Kluyveromyces Marxianus ◽  
Lipid Production ◽  
Carbon Sources ◽  
Industrial Applications ◽  
Human Society ◽  
Content Type ◽  
Renewable Chemicals

ABSTRACTThroughout history, the yeastSaccharomyces cerevisiaehas played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However,S. cerevisiaehas proven difficult to engineer to expand the carbon sources it can utilize, the products it can make, and the harsh conditions it can tolerate in industrial applications. Other yeasts that could solve many of these problems remain difficult to manipulate genetically. Here, we engineered the thermotolerant yeastKluyveromyces marxianusto create a new synthetic biology platform. Using CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, we show that wild isolates ofK. marxianuscan be made heterothallic for sexual crossing. By breeding two of these mating-type engineeredK. marxianusstrains, we combined three complex traits—thermotolerance, lipid production, and facile transformation with exogenous DNA—into a single host. The ability to crossK. marxianusstrains with relative ease, together with CRISPR-Cas9 genome editing, should enable engineering ofK. marxianusisolates with promising lipid production at temperatures far exceeding those of other fungi under development for industrial applications. These results establishK. marxianusas a synthetic biology platform comparable toS. cerevisiae, with naturally more robust traits that hold potential for the industrial production of renewable chemicals.IMPORTANCEThe yeastKluyveromyces marxianusgrows at high temperatures and on a wide range of carbon sources, making it a promising host for industrial biotechnology to produce renewable chemicals from plant biomass feedstocks. However, major genetic engineering limitations have kept this yeast from replacing the commonly used yeastSaccharomyces cerevisiaein industrial applications. Here, we describe genetic tools for genome editing and breedingK. marxianusstrains, which we use to create a new thermotolerant strain with promising fatty acid production. These results open the door to usingK. marxianusas a versatile synthetic biology platform organism for industrial applications.

scholarly journals Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae

2016 ◽  
Vol 127 ◽  
pp. 203-205 ◽  
Author(s):  
Wesley Cardoso Generoso ◽  
Manuela Gottardi ◽  
Mislav Oreb ◽  
Eckhard Boles
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Editing

scholarly journals Production of Recombinant Proteins in the Milk of Transgenic Animals: Current State and Prospects

Acta Naturae ◽  
2018 ◽  
Vol 10 (3) ◽  
pp. 40-47 ◽  
Author(s):  
M. V. Shepelev ◽  
S. V. Kalinichenko ◽  
A. V. Deykin ◽  
I. V. Korobko
Keyword(s):  
Genetic Engineering ◽  
Genome Editing ◽  
Recombinant Proteins ◽  
Retrospective Review ◽  
Current Trend ◽  
Transgenic Animals ◽  
Current State ◽  
The Creation ◽  
A Current

The use of transgenic animals as bioreactors for the synthesis of the recombinant proteins secreted into milk is a current trend in the development of biotechnologies. Advances in genetic engineering, in particular the emergence of targeted genome editing technologies, have provided new opportunities and significantly improved efficiency in the generation of animals that produce recombinant proteins in milk, including economically important animals. Here, we present a retrospective review of technologies for generating transgenic animals, with emphasis on the creation of animals that produce recombinant proteins in milk. The current state and prospects for the development of this area of biotechnology are discussed in relation to the emergence of novel genome editing technologies. Experimental and practical techniques are briefly discussed.

scholarly journals Genome Editing of Saccharomyces Cerevisiae Using CRISPR-Cas9 System

2021 ◽  
Vol 2 (1) ◽  
pp. 20-28
Author(s):  
Yaseen Ismael Imran ◽  
Ibrahim Abdulla Ahmed ◽  
Ahmed Ali Muhawesh
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Immune System ◽  
Genome Editing ◽  
Genetically Modified ◽  
Food Additives ◽  
Genetically Engineered ◽  
Flexible Tool ◽  
Cell Factories ◽  
Pharmaceutical Proteins ◽  
Long Time

Saccharomyces cerevisiae is an important yeast has been exploited for a long time to produce alcohol or bread. Moreover, genetically engineered S. cerevisiae cells continue to be used as cell factories for production of biofuels, pharmaceutical proteins and food additives. Genetically modified strain of S. cerevisiae created using traditional methods is laborious and time consuming. Recently, originally an immune system in archaea and bacteria, Clustered regularly interspaced short palindromic repeats “CRISPR” and CRISPR-associated “Cas” have been used exploited  as a flexible tool for genome editing. Until now, this tool has been applied to many organisms including yeast. Here, we review the importance of S. cerevisiae as an industrial platform and the use of CRISPR/Cas system and its applications in research and industry of this yeast.  

scholarly journals Simple-to-use CRISPR-SpCas9/SaCas9/AsCas12a vector series for genome editing in Saccharomyces cerevisiae

2021 ◽  
Author(s):  
Satoshi Okada ◽  
Goro Doi ◽  
Shitomi Nakagawa ◽  
Emiko Kusumoto ◽  
Takashi Ito
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Editing ◽  
Region Of Interest ◽  
Yeast Genome ◽  
Yeast Genetics ◽  
Reading Frame ◽  
Protospacer Adjacent Motif ◽  
Combined Use ◽  
Genome Manipulation ◽  
Complete Deletion

Genome editing using the CRISPR/Cas system has been implemented for various organisms and becomes increasingly popular even in the genetically tractable budding yeast Saccharomyces cerevisiae. Since each CRISPR/Cas system recognizes only the sequences flanked by its unique protospacer adjacent motif (PAM), a certain single system often fails to target a region of interest due to the lack of PAM, thus necessitating the use of another system with a different PAM. Three CRISPR/Cas systems with distinct PAMs, namely SpCas9, SaCas9, and AsCas12a, have been successfully used in yeast genome editing and their combined use should expand the repertoire of editable targets. However, currently available plasmids for these systems were individually developed under different design principles, thus hampering their seamless use in the practice of genome editing. Here we report a series of Golden Gate Assembly-compatible backbone vectors designed under a unified principle to exploit the three CRISPR/Cas systems in yeast genome editing. We also created a software to assist the design of genome-editing plasmids for individual target sequences using the backbone vectors. Genome editing with these plasmids demonstrated practically sufficient efficiency in both insertion of gene fragments to essential genes and complete deletion of an open reading frame. The backbone vectors with the software would thus provide a versatile toolbox to facilitate the seamless use of SpCas9, SaCas9, and AsCas12a in various types of genome manipulation, especially those that are difficult to perform with conventional techniques in yeast genetics.

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