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2015 ; 78
(ä): 99-107
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New technology and resources for cryptococcal research
#MMPMID25460849
Zhang N
; Park YD
; Williamson PR
Fungal Genet Biol
2015[May]; 78
(ä): 99-107
PMID25460849
show ga
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.