2026/03/23 by Haiting Wang, Yongjunlin Tan, Jiayue Ma +9 · 1 voice
Biochemistry, Genetics and Molecular Biology · Agricultural and Biological Sciences · #Genomics and Chromatin Dynamics #Fungal and yeast genetics research #Plant Molecular Biology Research
paper · doi:10.1002/mlf2.70060
openalex publication_date 2026/03/23 · openalex created_date 2026/03/25 · openalex updated_date 2026/07/23
Abstract Histone modifications and chromatin‐binding proteins play crucial roles in regulating gene expression in eukaryotes, with significant implications for fungal pathogenicity and development. However, profiling these modifications or proteins across the genome in fungi remains challenging due to the technical limitations of the traditional, widely used Chromatin Immunoprecipitation‐Sequencing (ChIP‐Seq) method. Here, we present an optimized fungal Cleavage Under Targets and Tagmentation‐Sequencing (fCUT&Tag‐Seq) protocol specifically designed for filamentous fungi and dimorphic fungi. Our approach involves the preparation of protoplasts and nuclear extraction to enhance antibody accessibility, along with formaldehyde crosslinking to improve protein‐DNA binding efficiency. We then successfully applied fCUT&Tag‐Seq to accurately profile multiple histone modifications like H3K9me3, H3K27me3, H3K4me3, and H3K18ac, across different plant pathogenic or model fungal species, including Verticillium dahliae , Neurospora crassa, Fusarium graminearum , and Sporisorium scitamineum , showing good signal‐to‐noise ratios, reproducibility, and detection sensitivity. Furthermore, we extended this method to profile chromatin‐binding proteins, such as the histone acetyltransferase Gcn5. This study establishes fCUT&Tag‐Seq as a robust and useful tool for fungal epigenetic research, enabling detailed exploration of chromatin dynamics and advancing our understanding of fungal gene regulation, development, and pathogenicity.