ArticlePloS one2016
Targeted Genome Editing via CRISPR in the Pathogen Cryptococcus neoformans.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 72 citations in OpenAlex.
- Advancements in CRISPR-Cas Systems for Genome Editing towards Eradication of Human Microbial Pathogens.Molecular biotechnology · 2026Review
- CRISPR/Cas9-compatible plasmids enabling seven dominant genetic selection methods for the human fungal pathogenMicrobiology spectrum · 2025Article
- Near 100% efficient homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans.G3 (Bethesda, Md.) · 2025Article
- Article
- Illuminating the Cryptococcus neoformans species complex: unveiling intracellular structures with fluorescent-protein-based markers.Genetics · 2024Article
- Improved Protoplast Production Protocol for Fungal Transformations Mediated by CRISPR/Cas9 inPlants (Basel, Switzerland) · 2024Article
- Biolistic Transformation of Cryptococcus neoformans.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Article
- Self-cleaving guide RNAs enable pharmacological selection of precise gene editing events in vivo.Nature communications · 2022Article
- Cryptococcus neoformans Database in Synthetic Biology Open Language.Microbiology resource announcements · 2022Article
- Genomic Approaches to Antifungal Drug Target Identification and Validation.Annual review of microbiology · 2022Review
- Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases.Journal of nanostructure in chemistry · 2022Review
- Ten decadal advances in fungal biology leading towards human well-being.Fungal diversity · 2022Review
- Homology length dictates the requirement for Rad51 and Rad52 in gene targeting in the Basidiomycota yeast Naganishia liquefaciens.Current genetics · 2021Article
- Hsp90 interaction networks in fungi-tools and techniques.FEMS yeast research · 2021Review
- Simplified All-In-One CRISPR-Cas9 Construction for Efficient Genome Editing inJournal of fungi (Basel, Switzerland) · 2021Article
- Recent Advances in Genome Editing Tools in Medical Mycology Research.Journal of fungi (Basel, Switzerland) · 2021Review
- Post-Translational Modifications Drive Success and Failure of Fungal-Host Interactions.Journal of fungi (Basel, Switzerland) · 2021Review
- Genetic Transformation inJournal of fungi (Basel, Switzerland) · 2021Review
- An intergenic "safe haven" region in Cryptococcus neoformans serotype D genomes.Fungal genetics and biology : FG & B · 2020Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Low rates of homologous integration have hindered molecular genetic studies in Cryptococcus neoformans over the past 20 years, and new tools that facilitate genome manipulation in this important pathogen are greatly needed. To this end, we have investigated the use of a Class 2 CRISPR system in C. neoformans (formerly C. neoformans var. grubii). We first expressed a derivative of the Streptococcus pyogenes Cas9 nuclease in C. neoformans, and showed that it has no effect on growth, production of virulence factors in vitro, or virulence in a murine inhalation model. In proof of principle experiments, we tested the CAS9 construct in combination with multiple self-cleaving guide RNAs targeting the well-characterized phosphoribosylaminoamidazole carboxylase-encoding ADE2 gene. Utilizing combinations of transient and stable expression of our constructs, we revealed that functionality of our CRISPR constructs in C. neoformans is dependent upon the CAS9 construct being stably integrated into the genome, whilst transient expression of the guide RNA is sufficient to enhance rates of homologous recombination in the CAS9 genetic background. Given that the presence of the CRISPR nuclease does not influence virulence in a murine inhalation model, we have successfully demonstrated that this system is compatible with studies of C. neoformans pathogenesis and represents a powerful tool that can be exploited by researchers in the field.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.