Abstract:
Candida viswanathii is a fungus that efficiently produces long-chain dicarboxylic
acids, which are useful in petrochemical production. It is closely related to Candida
tropicalis, a prominent human fungal pathogen. Because CRISPR has been
implemented in other Candida species, particularly Candida albicans, I hypothesized
that a similar CRISPR-mediated genome editing method could be developed for C.
viswanathii. I have sequenced the C. viswanathii genome and identified guide
sequences for ADE2 and CYP52. I then cloned these sequences into a vector that also
expresses the Cas9 nuclease. I cotransformed these vectors along with a repair
template that will introduce stop codons when incorporated into ADE2 and CYP52 of C.
viswanathii. I was able to obtain many transformants, and I am now determining if these
transformants have incorporated the repair template. My ultimate goal is to work in
collaboration to determine why and how C. viswanathii makes long-chain dicarboxylic
acids
