Development of ferret immune repertoire reference resources and single-cell-based high-throughput profiling assays

Area of Science:

• Immunology
• Genomics
• Comparative Medicine

Background:

• Domestic ferrets (Mustela putorius furo) are valuable models for human respiratory diseases.
• Limited identification and annotation of ferret B and T cell receptors hinder immune repertoire studies.
• Lack of ferret-specific reagents impedes detailed analysis of their immune system.

Purpose of the Study:

• To create a comprehensive reference set of ferret immunoglobulin (Ig) and T cell receptor (TCR) genes.
• To develop and validate novel ferret-specific immune repertoire profiling assays.
• To enhance the study of ferret immune responses and their application in disease modeling.

Main Methods:

• Long-read transcriptome sequencing of ferret splenocyte and lymph node samples.
• Systematic annotation of germline Ig and TCR variable (V), diversity (D), joining (J), and constant (C) genes.
• Design and experimental validation of ferret-specific immune repertoire profiling assays using single-cell technology.

Main Results:

• Obtained over 120,000 high-quality full-length Ig and TCR transcripts.
• Constructed a complete reference set for ferret Ig and TCR constant regions.
• Developed and validated novel ferret-specific assays compatible with single-cell platforms.

Conclusions:

• The developed resources and assays provide a foundation for comprehensive ferret immune repertoire studies.
• These advancements will enable more accurate analyses of ferret immune diversity.
• Improved tools will facilitate the use of ferrets as models for human respiratory diseases.