A genome-scale resource for in vivo tag-based protein function exploration in C. elegans

Area of Science:

• Genetics and Genomics
• Molecular Biology
• Developmental Biology

Background:

• Understanding protein localization and interactions in vivo is crucial for deciphering biological processes.
• Existing methods often lack the scale and precision needed for comprehensive proteome analysis in a multicellular organism.

Purpose of the Study:

• To develop a genome-scale transgenic platform for in vivo protein studies in Caenorhabditis elegans.
• To create a comprehensive resource for systematic protein function interrogation under endogenous regulatory control.

Main Methods:

• Genome-scale transgenic platform construction using computer-assisted design, massively parallel DNA engineering, and next-generation sequencing.
• Generation of 14,637 genomic DNA transgenes covering 73% of the C. elegans proteome.
• Utilized a multipurpose tag for in vivo protein localization and affinity purification.

Main Results:

• Successfully generated a resource of 14,637 genomic DNA transgenes for C. elegans.
• Demonstrated the platform's utility through systematic chromatin immunopurification and automated 4D imaging.
• Produced detailed DNA binding and cell/tissue distribution maps for key transcription factors.

Conclusions:

• The developed platform provides a powerful tool for large-scale in vivo protein studies in a multicellular organism.
• This resource enables detailed mapping of protein functions, including DNA binding and localization patterns.
• Facilitates systematic interrogation of protein function and gene regulation in C. elegans.