SWITCH: a dynamic CRISPR tool for genome engineering and metabolic pathway control for cell factory construction in Saccharomyces cerevisiae.
Microb Cell Fact
; 16(1): 25, 2017 Feb 08.
Article
en En
| MEDLINE
| ID: mdl-28179021
ABSTRACT
BACKGROUND:
The yeast Saccharomyces cerevisiae is increasingly used as a cell factory. However, cell factory construction time is a major obstacle towards using yeast for bio-production. Hence, tools to speed up cell factory construction are desirable.RESULTS:
In this study, we have developed a new Cas9/dCas9 based system, SWITCH, which allows Saccharomyces cerevisiae strains to iteratively alternate between a genetic engineering state and a pathway control state. Since Cas9 induced recombination events are crucial for SWITCH efficiency, we first developed a technique TAPE, which we have successfully used to address protospacer efficiency. As proof of concept of the use of SWITCH in cell factory construction, we have exploited the genetic engineering state of a SWITCH strain to insert the five genes necessary for naringenin production. Next, the naringenin cell factory was switched to the pathway control state where production was optimized by downregulating an essential gene TSC13, hence, reducing formation of a byproduct.CONCLUSIONS:
We have successfully integrated two CRISPR tools, one for genetic engineering and one for pathway control, into one system and successfully used it for cell factory construction.Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Saccharomyces cerevisiae
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Ingeniería Genética
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Redes y Vías Metabólicas
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Sistemas CRISPR-Cas
Idioma:
En
Revista:
Microb Cell Fact
Asunto de la revista:
BIOTECNOLOGIA
/
MICROBIOLOGIA
Año:
2017
Tipo del documento:
Article
País de afiliación:
Dinamarca