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Protein interaction networks and subcellular proteomes can provide key insights into the biological functions of protein complexes. Here, we describe a method for proximity-based labeling using TurboID to define protein interaction networks in Cryptosporidium parvum. In addition, we have developed an auxin-inducible degron (AID) labeling system for conditional protein degradation in C. parvum species to facilitate functional studies. This chapter describes the procedures for generating miniTurbo or mAID-TIR1-tagged C. parvum species strains and validating the TurboID and mAID systems for target genes.

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Establishing TurboID Proximity Labeling and mAID Conditional Knockdown System in Cryptosporidium parvum

  • 实验方案

  • 发表日期：

    2026年

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  • Rui Xu,
  • L. David Sibley

摘要

Protein interaction networks and subcellular proteomes can provide key insights into the biological functions of protein complexes. Here, we describe a method for proximity-based labeling using TurboID to define protein interaction networks in Cryptosporidium parvum. In addition, we have developed an auxin-inducible degron (AID) labeling system for conditional protein degradation in C. parvum species to facilitate functional studies. This chapter describes the procedures for generating miniTurbo or mAID-TIR1-tagged C. parvum species strains and validating the TurboID and mAID systems for target genes.