title: New technology and resources for cryptococcal research.
abstract: Rapid advances in molecular biology and genome sequencing have enabled the generation of new technology and resources for cryptococcal research. RNAi-mediated specific gene knock down has become routine and more efficient by utilizing modified shRNA plasmids and convergent promoter RNAi constructs. This system was recently applied in a high-throughput screen to identify genes involved in host-pathogen interactions. Gene deletion efficiencies have also been improved by increasing rates of homologous recombination through a number of approaches, including a combination of double-joint PCR with split-marker transformation, the use of dominant selectable markers and the introduction of Cre-Loxp systems into Cryptococcus. Moreover, visualization of cryptococcal proteins has become more facile using fusions with codon-optimized fluorescent tags, such as green or red fluorescent proteins or, mCherry. Using recent genome-wide analytical tools, new transcriptional factors and regulatory proteins have been identified in novel virulence-related signaling pathways by employing microarray analysis, RNA-sequencing and proteomic analysis.
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institution of Health, Bethesda, MD, United States.
authors.forename: Nannan
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institution of Health, Bethesda, MD, United States.
authors.forename: Yoon-Dong
authors.initials: YD
authors.lastname: Park
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institution of Health, Bethesda, MD, United States. Electronic address: williamsonpr@mail.nih.gov.
authors.forename: Peter R
authors.initials: PR
authors.lastname: Williamson
mesh_terms: D035843:Biomedical Research / Q000379:methods* / Q000639:trends; D003453:Cryptococcosis / Q000382:microbiology*; D003454:Cryptococcus / Q000235:genetics* / Q000472:pathogenicity*; D020869:Gene Expression Profiling / Q000379:methods; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods; D023281:Genomics / Q000379:methods; D054884:Host-Pathogen Interactions; D008828:Microbiological Techniques / Q000379:methods* / Q000639:trends; D008967:Molecular Biology / Q000379:methods* / Q000639:trends; D040901:Proteomics / Q000379:methods
keywords: Cryptococcosis; Cryptococcus gattii; Cryptococcus neoformans; Pathogenic fungus; RNAi; Yeast