The
emergence of the new coronavirus 2019 (COVID-19) was first
seen in December 2019, which has spread rapidly and become a global
pandemic. The number of cases of COVID-19 and its associated mortality
have raised serious concerns worldwide. Early diagnosis of viral infection
undoubtedly allows rapid intervention, disease management, and substantial
control of the rapid spread of the disease. Currently, the standard
approach for COVID-19 diagnosis globally is the RT-qPCR test; however,
the limited access to kits and associated reagents, the need for specialized
lab equipment, and the need for highly skilled personnel has led to
a detection slowdown. Recently, the development of clustered regularly
interspaced short palindromic repeats (CRISPR)-based diagnostic systems
has reshaped molecular diagnosis. The benefits of the CRISPR system
such as speed, precision, specificity, strength, efficiency, and versatility
have inspired researchers to develop CRISPR-based diagnostic and therapeutic
methods. With the global COVID-19 outbreak, different groups have
begun to design and develop diagnostic and therapeutic programs based
on the efficient CRISPR system. CRISPR-based COVID-19 diagnostic systems
have advantages such as a high detection speed (i.e., 30 min from
raw sample to reach a result), high sensitivity and precision, portability,
and no need for specialized laboratory equipment. Here, we review
contemporary studies on the detection of COVID-19 based on the CRISPR
system.
Manipulating fungal genomes is an important tool to understand the function of target genes, pathobiology of fungal infections, virulence potential, and pathogenicity of medically important fungi, and to develop novel diagnostics and therapeutic targets. Here, we provide an overview of recent advances in genetic manipulation techniques used in the field of medical mycology. Fungi use several strategies to cope with stress and adapt themselves against environmental effectors. For instance, mutations in the 14 alpha-demethylase gene may result in azole resistance in Aspergillusfumigatus strains and shield them against fungicide’s effects. Over the past few decades, several genome editing methods have been introduced for genetic manipulations in pathogenic fungi. Application of restriction enzymes to target and cut a double-stranded DNA in a pre-defined sequence was the first technique used for cloning in Aspergillus and Candida. Genome editing technologies, including zinc-finger nucleases (ZFNs) and transcriptional activator-like effector nucleases (TALENs), have been also used to engineer a double-stranded DNA molecule. As a result, TALENs were considered more practical to identify single nucleotide polymorphisms. Recently, Class 2 type II Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9 technology has emerged as a more useful tool for genome manipulation in fungal research.
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