Jun 29, 2017

Development of a CRISPR-Cas9 System for Efficient Genome Editing of Candida lusitaniae

MSphere
Emily L NortonRichard J Bennett

Abstract

Candida lusitaniae is a member of the Candida clade that includes a diverse group of fungal species relevant to both human health and biotechnology. This species exhibits a full sexual cycle to undergo interconversion between haploid and diploid forms. C. lusitaniae is also an emerging opportunistic pathogen that can cause serious bloodstream infections in the clinic and yet has often proven to be refractory to facile genetic manipulations. In this work, we develop a clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated gene 9 (Cas9) system to enable genome editing of C. lusitaniae. We demonstrate that expression of CRISPR-Cas9 components under species-specific promoters is necessary for efficient gene targeting and can be successfully applied to multiple genes in both haploid and diploid isolates. Gene deletion efficiencies with CRISPR-Cas9 were further enhanced in C. lusitaniae strains lacking the established nonhomologous end joining (NHEJ) factors Ku70 and DNA ligase 4. These results indicate that NHEJ plays an important role in directing the repair of DNA double-strand breaks (DSBs) in C. lusitaniae and that removal of this pathway increases integration of gene deletion templates by homolo...Continue Reading

  • References31
  • Citations4

References

Mentioned in this Paper

Study
Non-Homologous DNA End-Joining
Biochemical Pathway
Clinic
DNA Repair
Genes
CRISPR-Cas Systems
Pathogenic Organism
Recombination, Genetic
Gene Deletion Abnormality

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