2024-03-28T18:56:41Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1474072017-04-24T07:20:43Zcom_10261_10735com_10261_2col_10261_10737
Vigentini, Ileana
Gebbia, Marinella
Zhou, Shijie
Foschino, Roberto
Morales, Pilar
González García, Ramón
Roth, Frederick P.
2017-03-27T11:45:34Z
2017-03-27T11:45:34Z
2015-10
3rd International Conference on Microbial Diversity (2015)
http://hdl.handle.net/10261/147407
Saccharomyces cerevisiae has an extensive repertoire of molecular methods for the genetic
analysis of laboratory strains. Although this has widely contributed to the interpretation of
gene functionality within haploid genomes, the genetic basis of desirable metabolic pathways
in industrial diploid and polyploid yeast strain is still poorly understood. The genetic
engineering of industrial yeasts is currently undergoing major changes due to the
development of a marker-free, high-throughput, and multiplexed genome editing approach
inspired to the bacterial immune systems: the “Clustered Regularly Interspaced Short
Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) immune system”.
CRISPR/Cas9 allows the knockout or the tuning of expression of specific targeted genes and
pathways. The system requires an endonuclease (Cas9) that is guided to a specific DNA
sequence by a guide RNA (gRNA). The assembling of a ribonucleoprotein complex
(Cas9/gRNA) can produce cuts in specific sites of eukaryotic genomes and generates
different combinations of gene deletions through a single transformation event.
CRISPR/Cas9 is gaining significant attention because of its biotechnological applications
and its potential to boost innovation. The wine industry could particularly gain an advantage
from this engineering system; indeed, molecular studies should help understanding the
contribution of Saccharomyces and non-Saccharomyces species to several wine features such
as those linked to wine quality and safety (aroma and off-flavours compounds, ethanol and
glycerol production, sulphur dioxide resistance, toxic compound formation, etc.). In this
study we outline a strategy to modify wine yeasts by CRISPR/Cas9 approach. To prove the
robustness of this system and to offers an engineering pipeline for further gene editing in
specific metabolic pathways relevant for the wine production, two commercial strains of
S. cerevisiae (EC1118 and AWRI 796) have been genetically engineered in the arginine
degradation pathway to generate strains with a reduced urea and ethyl-carbamate production.
eng
closedAccess
CRISPR/CAS9 system as a valuable genome editing tool applicate to wine yeasts
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