Antifungal Mechanism of Action (MoA) Study Service

Primary MoA Screens

• Cytological Profiling (FCP): A rapid and powerful phenotypic screen using fluorescent dyes to visualize the effect of your compound on various cellular compartments (e.g., cell wall, cell membrane, nucleus, mitochondria) in real-time. This provides an initial "fingerprint" of the MoA.
• Phenotypic Rescue Assays: Using a library of fungal strains with defined defects (e.g., auxotrophs), we can test if your compound's toxicity can be "rescued" by supplementing the growth medium with specific nutrients. This can indicate interference with a metabolic pathway, such as amino acid or nucleic acid synthesis.

Genetic & Molecular Approaches

• Mutant Strain Profiling: We screen your compound against our extensive biobank of characterized fungal strains, including those with mutations in key antifungal target genes (e.g., ERG11 for azoles, FKS1 for echinocandins). This helps determine if your compound shares a target with existing drugs or acts via a novel pathway.
• Gene Deletion and Overexpression Libraries: We can utilize these powerful tools in model organisms like Saccharomyces cerevisiae to identify genes whose deletion or overexpression confers resistance or sensitivity to your compound. This provides a direct link between your drug and its genetic target.
• Target Deconvolution via "-omics" Technologies
  • Transcriptomics (RNA-Seq): We compare gene expression profiles of treated and untreated fungal cells to identify which genes are upregulated or downregulated in response to your compound. The resulting "transcriptional signature" can be matched to known pathways, providing strong evidence for a particular MoA.
  • Proteomics: We analyze the entire protein complement of the cell to see which proteins are affected by your compound.
  • Chemical Proteomics: We can use affinity-based probes to directly "pull down" and identify the protein(s) that your compound binds to.

Biochemical & Biophysical Assays

• Enzyme Inhibition Assays: If transcriptomics or genetic screens point to a specific enzyme target (e.g., a demethylase, a synthase), we can develop and perform in vitro assays to confirm direct enzyme inhibition and determine key kinetic parameters.
• Cell Membrane and Cell Wall Integrity Assays: Using specialized dyes and assays, we can quantify the effects of your compound on the integrity and permeability of the fungal cell wall and membrane.
• Time-Kill Kinetics: We determine the rate at which your compound kills fungal cells, providing insight into whether it is fungicidal (killing the fungus) or fungistatic (inhibiting its growth).

Sample Information

We typically require 1-5 mg of your compound, with purity and solubility information.

Deliverables

A comprehensive final report including:

• Summary of findings and key conclusions
• Detailed methodology for each assay
• Raw data and analyzed results (e.g., graphs, tables, heatmaps)
• Interpretation of MoA and recommendations for future studies
• High-resolution images from microscopy and other imaging techniques

Turnaround Time

Project timelines vary depending on the scope and complexity of the services requested. Preliminary MoA screens can be completed in as little as 4-6 weeks, while a full, integrated study with "-omics" profiling may take 3-6 months. We will provide a detailed timeline in your custom project proposal.