Authors: Elvira Román (Complutense University of Madrid, Spain)
Take a look behind the scenes of a recent Future Microbiology article, entitled ‘New insights of CRISPR technology in pathogenic fungi‘, as we ask author Elvira Román (Complutense University of Madrid, Spain) about the advances CRISPR technology has already brought to this field and the potential it could hold in the future.
What inspired you to write this piece?
If you are researcher in biology and make use of genetics, then CRISPR is an amazing tool. Several years ago, when I started my PhD doing a deletion in Candida albicans, our favourite organism, while feasible, was a tedious and time-consuming task. However, it is now easy to test biological hypothesis on gene function. Disseminating this technology will contribute to its popularization which will be important in our field of research.
What makes CRISPR technology so useful for studying human pathogenic fungi?
The main advantage of CRISPR is the specificity and efficiency of this genome-editing technology, which is based on Watson–Crick base pairing. This is much simpler than the available transcription activator-like effector nucleases (TALENs) or zinc-finger nucleases (ZFNs) for edition of genomes. More importantly, it can be accomplished in a short period of time and is comparatively cheap. It allows simultaneous and efficient editing of several genes, which is essential for diploid fungi, making easy to develop strains suitable for phenotype characterization, such as those altered in putative virulence traits, a fact with potential impact in human health.
What progresses have been made in using CRISPR technology in human pathogenic fungi?
Up to now, CRISPR has been completely implemented in a wide range of fungi, including filamentous fungi with an important role in biotechnology. The possibility of modifying one, or several, genes simultaneously in really short periods of time and in a very efficient manner has served to overcome the lack of a readily available genetic toolkits for non-conventional model organisms, including fungi.
What areas of CRISPR technology could be optimised further for use in this field?
Control of gene expression using catalytically inactive versions of Cas9 can be used to either increase or repress genes. Most importantly, a set of genes could be simultaneously activated while another set repressed, enabling new possibilities in the analysis of complex metabolic pathways involved in the production of certain metabolites, virulence or resistance to antifungal drugs. In pathogenic fungi, this strategy can be used to circumvent problems associated with essential genes.
What are your predictions for next 10–15 years in this field? What do you hope to see?
CRISPR will be a routine tool for gene editing in many biological systems. The unique properties of Cas9 enzyme as a structural scaffold for the interaction of the three main components of life (RNA, DNA and proteins) will enable several possibilities in research. New and more creative tools will be developed and they will be expanded with the use of additional CRISPR systems (not only streptococcal Cas9).
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