Simplified All-In-One CRISPR-Cas9 Construction for Efficient Genome Editing in Cryptococcus Species.

ABSTRACT: Cryptococcus neoformans and Cryptococcus deneoformans are opportunistic fungal pathogens found worldwide that are utilized to reveal mechanisms of fungal pathogenesis. However, their low homologous recombination frequency has greatly encumbered genetic studies. In preliminary work, we described a 'suicide' CRISPR-Cas9 system for use in the efficient gene editing of C. deneoformans, but this has not yet been used in the C. neoformans strain. The procedures involved in constructing vectors are time-consuming, whether they involve restriction enzyme-based cloning of donor DNA or the introduction of a target sequence into the gRNA expression cassette via overlap PCR, as are sophisticated, thus impeding their widespread application. Here, we report the optimized and

SUBMITTER: Zhang P 

PROVIDER: S-EPMC8303259 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature