ArticleNature protocols2019
Design, execution, and analysis of CRISPR-Cas9-based deletions and genetic interaction networks in the fungal pathogen Candida albicans.
Article in Nature protocols, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Pooled CRISPRi screening reveals fungal-specific drug target candidates.Nature microbiology · 2026Article
- The unique Efg1 fungal virulence regulon in the catheterized bladder environment.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Hac1-independent functions of Ire1 drive morphogenesis in Candida albicans through distinct transcriptional programs.Journal of cell science · 2026Article
- Advancements in CRISPR-Cas Systems for Genome Editing towards Eradication of Human Microbial Pathogens.Molecular biotechnology · 2026Review
- EditingBiomolecules · 2026Review
- Pooled CRISPRi screening reveals fungal-specific vulnerabilities across environments and genetic backgrounds.bioRxiv : the preprint server for biology · 2026Article
- Article
- PMA1-containing extracellular vesicles ofGut microbes · 2025Article
- The putative error prone polymerasenpj antimicrobials and resistance · 2024Article
- Updated immunomodulatory roles of gut flora and microRNAs in inflammatory bowel diseases.Clinical and experimental medicine · 2023Review
- The catheterized bladder environment promotes Efg1- and Als1-dependentScience advances · 2023Article
- Development and applications of a CRISPR activation system for facile genetic overexpression in Candida albicans.G3 (Bethesda, Md.) · 2023Article
- Ten decadal advances in fungal biology leading towards human well-being.Fungal diversity · 2022Review
- The SAGA and NuA4 component Tra1 regulates Candida albicans drug resistance and pathogenesis.Genetics · 2021Article
- Recent Advances in Genome Editing Tools in Medical Mycology Research.Journal of fungi (Basel, Switzerland) · 2021Review
- Comprehensive genetic analysis of adhesin proteins and their role in virulence of Candida albicans.Genetics · 2021Article
- Combining Genome-Wide Gene Expression Analysis (RNA-seq) and a Gene Editing Platform (CRISPR-Cas9) to Uncover the Selectively Pro-oxidant Activity of Aurone Compounds AgainstFrontiers in microbiology · 2021Article
- Advances in Molecular Tools and In Vivo Models for the Study of Human Fungal Pathogenesis.Microorganisms · 2020Review
- The CRISPR toolbox in medical mycology: State of the art and perspectives.PLoS pathogens · 2020Review
- CRISPR-Based Genetic Manipulation ofFrontiers in genome editing · 2020Review
Corrections and comments
- Erratum issued
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The study of fungal pathogens is of immediate importance, yet progress is hindered by the technical challenges of genetic manipulation. For Candida species, their inability to maintain plasmids, unusual codon usage, and inefficient homologous recombination are among the obstacles limiting efficient genetic manipulation. New advances in genomic biotechnologies-particularly CRISPR-based tools-have revolutionized genome editing for many fungal species. Here, we present a protocol for CRISPR-Cas9-based manipulation in Candida albicans using a modified gene-drive-based strategy that takes ~1 month to complete. We detail the generation of Candida-optimized Cas9-based plasmids for gene deletion, an efficient transformation protocol using C. albicans haploids, and an optimized mating strategy to generate homozygous single- and double-gene diploid mutants. We further describe protocols for quantifying cell growth and analysis pipelines to calculate fitness and genetic interaction scores for genetic mutants. This protocol overcomes previous limitations associated with genetic manipulation in C. albicans and advances researchers' ability to perform genetic analysis in this pathogen; the protocol also has broad applicability to other mating-competent microorganisms.
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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.