CRISPR-cas system for Clostridium genome engineering and recombinant strains produced thereof
AssigneeAuburn University
InventorsYi Wang, Jie Zhang
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Abstract
A system for modifying the genome of Clostridium strains is provided based on a modified endogenous CRISPR array. The application also describes Clostridium strains modified for enhanced butanol production wherein the modified strains are produced using the novel CRISPR-Cas system.
Claims11
A Clostridium strain modified for enhanced butanol production relative to wild type C. tyrobutyricum (ATCC 25755), said Clostridium strain comprising
- a modification to the native cat1 gene, said modification preventing expression of a functional cat1 gene product;
- and
- an exogenous sequence encoding
- i) an aldehyde dehydrogenase;
- ii) a bifunctional aldehyde/alcohol dehydrogenase;
- or
- iii) an aldehyde dehydrogenase and an alcohol dehydrogenase.
A Clostridium strain modified for enhanced butanol production relative to wild type C. tyrobutyricum (ATCC 25755), said Clostridium strain being selected from the group consisting of Clostridium butyricum, Clostridium thermobutyricum, Clostridium cellulovorans, Clostridium carboxidivorans, Clostridium tyrobutyricum, Clostridium polysaccharolyticum, Clostridium populeti, and Clostridium kluyveri and comprising
- a modification to the native cat1 gene, said modification preventing expression of a functional cat1 gene product;
- and
- an adhE gene introduced into said cell and having at least 95% sequence identity to a C. acetobutylicum aldehyde/alcohol dehydrogenase gene of SEQ ID NO:
- 133 or SEQ ID NO:
- 134.
Dependencies are inferred from the claim text.