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HUH endonuclease-mediated DNA-protein conjugates: sequence-specific tools and cellular applications
Xiaowei Huang1,2, Haokun Li2, Jiajun Du2
1School of Chemistry and Chemical Engineering, University of South China, Hengyang, 421001, China.
Chemical Communications (Cambridge, England)
|October 1, 2025
Summary
HUH endonucleases act as self-labeling protein tags for DNA-protein conjugation. These tools enable applications in cellular engineering, therapeutics, and genome editing, with future potential in RNA targeting.
Area of Science:
- Chemical Biology
- Synthetic Biology
- Biotechnology
Background:
- HUH endonucleases are enzymes with conserved HUH and Y motifs.
- These motifs facilitate sequence-specific covalent conjugation of single-stranded DNA (ssDNA).
- Their bioorthogonal and robust nature allows conjugation under physiological conditions.
Purpose of the Study:
- To review recent advancements in using HUH endonucleases as self-labeling protein tags.
- To examine the applications of engineered DNA-protein conjugates in cellular studies.
- To highlight future directions in HUH endonuclease research.
Main Methods:
- Structural and catalytic mechanism analysis of HUH and Y motifs.
- Review of recent applications of HUH-based DNA-protein conjugates.
- Exploration of computational tools and directed evolution strategies.
Main Results:
- HUH endonucleases enable selective and bioorthogonal conjugation of ssDNA to proteins.
- Engineered DNA-protein conjugates show versatility in cellular interface engineering, drug delivery, and CRISPR-Cas systems.
- Emerging directions include computational design and RNA substrate targeting.
Conclusions:
- HUH endonucleases are powerful tools for creating DNA-protein conjugates.
- These conjugates drive innovation in chemical biology, synthetic biology, and therapeutics.
- Future research focuses on computational design and expanding HUH reactivity to RNA.
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