ArticleNature protocols2020
Genome-wide piggyBac transposon-based mutagenesis and quantitative insertion-site analysis in haploid Candida species.
Article in Nature protocols, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 15 citations in OpenAlex.
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- Genome-Wide Landscape of Position Effect Variegation inJournal of fungi (Basel, Switzerland) · 2026Article
- Production of triacetic acid lactone from oleic acid by engineering the yeast Candida viswanathii.Microbial cell factories · 2025Article
- Signaling pathways governing the pathobiological features and antifungal drug resistance ofmBio · 2025Review
- Sdd3 regulates the biofilm formation ofmBio · 2025Article
- Engineering immunity with CAR-NK cells: advancing the frontiers of cancer immunotherapy.Frontiers in pharmacology · 2025Review
- High-throughput and genome-scale targeted mutagenesis using CRISPR in a nonmodel multicellular organism,Genome research · 2024Article
- Fungal resilience and host-pathogen interactions: Future perspectives and opportunities.Parasite immunology · 2023Review
- Inactivating the mannose-ethanolamine phosphotransferase Gpi7 confers caspofungin resistance in the human fungal pathogenCell surface (Amsterdam, Netherlands) · 2021Article
- LncRNA DINOR is a virulence factor and global regulator of stress responses in Candida auris.Nature microbiology · 2021Article
- CAR-NK Cell: A New Paradigm in Tumor Immunotherapy.Frontiers in oncology · 2021Review
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Invasive fungal infections caused by Candida species are life threatening with high mortality, posing a severe public health threat. New technologies for rapid, genome-wide identification of virulence genes and therapeutic targets are urgently needed. Our recent engineering of a piggyBac (PB) transposon-mediated mutagenesis system in haploid Candida albicans provides a powerful discovery tool, which we anticipate should be adaptable to other haploid Candida species. In this protocol, we use haploid C. albicans as an example to present an improved version of the mutagenesis system and provide a detailed description of the protocol for constructing high-quality mutant libraries. We also describe a method for quantitative PB insertion site sequencing, PBISeq. The PBISeq library preparation procedure exploits tagmentation to quickly and efficiently construct sequencing libraries. Finally, we present a pipeline to analyze PB insertion sites in a de novo assembled genome of our engineered haploid C. albicans strain. The entire protocol takes ~7 d from transposition induction to having a final library ready for sequencing. This protocol is highly efficient and less labor intensive than alternative approaches and significantly accelerates genetic studies of Candida.
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