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CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
Published on: June 9, 2020
Recent Advances in Genome Editing Tools in Medical Mycology Research
Sanaz Nargesi1, Saeed Kaboli2,3, Jose Thekkiniath4
1Department of Medical Mycology, School of Medicine, Mazandaran University of Medical Sciences, Sari 481751665, Iran.
Abstract:
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.
Insights
Genetic manipulation techniques are crucial for understanding fungal infections and developing new treatments. Recent advances, including CRISPR/Cas9, offer powerful tools for medical mycology research.
Area of Science:
- Medical Mycology
- Genetics
- Molecular Biology
Background:
- Fungal infections pose significant health risks, necessitating research into their pathogenicity and virulence.
- Understanding fungal gene function is key to developing diagnostics and therapeutics.
- Fungi exhibit adaptive strategies, such as azole resistance in Aspergillus fumigatus, impacting treatment efficacy.
Purpose of the Study:
- To provide an overview of recent advancements in genetic manipulation techniques for medically important fungi.
- To highlight the evolution of genome editing tools in medical mycology.
- To emphasize the utility of these techniques in studying fungal pathobiology and developing novel interventions.
Main Methods:
- Review of historical and recent genetic manipulation techniques.
- Discussion of restriction enzymes for DNA manipulation.
- Exploration of zinc-finger nucleases (ZFNs) and transcriptional activator-like effector nucleases (TALENs).
- Focus on the application of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9 technology.
Main Results:
- Restriction enzymes were early tools for cloning in Aspergillus and Candida.
- ZFNs and TALENs enabled targeted DNA modification, with TALENs proving useful for single nucleotide polymorphism identification.
- CRISPR/Cas9 has emerged as a highly effective and versatile tool for fungal genome manipulation.
Conclusions:
- Genetic manipulation is indispensable for advancing medical mycology.
- The progression of genome editing technologies, particularly CRISPR/Cas9, has significantly enhanced research capabilities.
- These tools are vital for dissecting fungal virulence, resistance mechanisms, and for the development of targeted antifungal strategies.
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