A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans

Area of Science:

• Developmental Biology
• Molecular Biology
• Genetics

Background:

• Proteins crucial for early embryonic development are often reused in later stages and in adults.
• Understanding protein function across different biological contexts necessitates tissue-specific experimental approaches.
• Existing methods for studying gene function may lack the precision for tissue-specific analysis.

Purpose of the Study:

• To develop a novel, efficient method for tissue-specific protein ablation in C. elegans.
• To enable the study of protein function in distinct tissues and developmental stages.
• To provide a versatile tool for C. elegans research.

Main Methods:

• Utilized CRISPR-Cas9 for endogenous GFP tagging of proteins.
• Developed a transgene expressing a fusion of a GFP-targeting nanobody and a SOCS-box ubiquitin ligase adaptor.
• Combined GFP tagging with the nanobody-ligase system for targeted protein degradation in specific tissues.

Main Results:

• Successfully demonstrated tissue-specific protein ablation in multiple C. elegans tissues, including epidermis, intestine, muscle, and neurons.
• The method recapitulated known loss-of-function phenotypes, validating its efficacy.
• The approach proved efficient and routine for targeted protein degradation.

Conclusions:

• The developed nanobody-ligase system provides a powerful and versatile tool for C. elegans research.
• This method facilitates the routine analysis of tissue-specific protein functions.
• The toolkit and strains will complement existing approaches, advancing the study of gene function in vivo.