RESEARCHERS DEVELOP CRISPR-CAS9 EDITING TECHNIQUES FOR RALSTONIA EUTROPHA
Ralstonia eutropha is an important bacterium for the study of polyhydroxyalkanoate (PHA) synthesis and CO2 fixation, making it a potential strain for industrial PHA production and CO2 conversion. However, current methods for genome editing the microbe are inefficient. Thus, developing an efficient method for R. eutropha genome editing is imperative.
The team of Bin Xiong from the University of Chinese Academy of Sciences developed a method for R. eutropha using an electroporation-based CRISPR-Cas9 technique. Using the new method, five R. eutropha genes were successfully edited via homologous recombination, with efficiencies ranging from 78.3 to 100%. The team also found that fructose can reduce the expression of the arabinose-inducible pBAD promoter, which was used to optimize the expression of cas9.
The CRISPR-Cpf1 system and the non-homologous end joining mechanism were also investigated, but failed to yield edited strains.
This study presents the first genome editing method for R. eutropha using an electroporation-based CRISPR-Cas9 approach. This will facilitate more studies and applications of R. eutrophafor PHA production and CO2 conversion.
For more information, read the article in Biotechnology for Biofuels.