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Genome editing of using CRISPR/Cas9

Biotechnol Biofuels. 2016-10-01; 
Shilpa Nagaraju, Naomi Kathleen Davies, David Jeffrey Fraser Walker, Michael Köpke, Séan Dennis Simpson
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Codon Optimization … The cas9 gene was codon adapted to C. autoethanogenum using GenScript's algorithm and C. autoethanogenum codon frequency table. The codon-adapted cas9 gene (GenBank: KU867916) was synthesized by GenScript Get A Quote

摘要

background: Impactful greenhouse gas emissions abatement can now be achieved through gas fermentation using acetogenic microbes for the production of low-carbon fuels and chemicals. However, compared to traditional hosts like or yeast, only basic genetic tools exist for gas-fermenting acetogens. To advance the process, a robust genetic engineering platform for acetogens is essential. results: In this study, we report scarless genome editing of an industrially used model acetogen, , using the CRISPR/Cas9 system. Initial efforts to retrofit the CRISPR/Cas9 system for resulted in poor efficiency likely due to uncontrolled expression of Cas9. To address this, we constructed and screened a small library of tetrac... More

关键词

Acetogen, CRISPR/Cas9, Clostridium autoethanogenum, Gas fermentation, Genome editing, Inducible promoter
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