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Engineered hypercompact Fanzor-ωRNA system with enhanced genome editing activity.

Guoli Zhao1, Yuling Liu1, Ganggang Zhang1

  • 1Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Key Laboratory of Myopia and Related Eye Diseases, Key Laboratory of Myopia and Related Eye Diseases, Chinese Academy of Medical Sciences, Shanghai 200031, China; Shanghai Research Center of Ophthalmology and Optometry, Eye & ENT Hospital, Fudan University, Shanghai 200031, China.

Molecular Cell
|October 10, 2025
PubMed
Summary

Engineered Fanzor nucleases show high genome editing activity in humans. This compact SpuFz1 V4 system enables efficient in vivo editing, advancing gene therapy potential.

Keywords:
FanzorRNA-guided DNA nucleasesSpuFz1base editinggenome editingin vivo retinal editingprotein engineeringsingle-AAV delivery

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Fanzor proteins are RNA-guided DNA nucleases found in eukaryotes.
  • Their compact size offers potential for genome editing applications.
  • Existing Fanzor systems exhibit low editing efficiency in mammalian genomes.

Purpose of the Study:

  • To engineer a Fanzor-based DNA endonuclease with enhanced editing activity.
  • To evaluate the potential of the engineered Fanzor as a base editor.
  • To demonstrate in vivo genome editing applications using the novel Fanzor system.

Main Methods:

  • Engineering of SpuFz1 V4, an RNA-guided DNA endonuclease.
  • Assessing editing efficiency in the human genome.
  • Delivery of SpuFz1 V4 via adeno-associated virus (AAV) for in vivo studies.

Main Results:

  • SpuFz1 V4 exhibits vigorous editing activity in the human genome, surpassing previous Fanzor nucleases.
  • The Fanzor system demonstrates substantial potential as a base editor.
  • Efficient in vivo genome editing was achieved in the retina using AAV delivery.

Conclusions:

  • SpuFz1 V4 is a highly active eukaryotic RNA-guided DNA nuclease.
  • The compact SpuFz1 V4 system facilitates efficient in vivo genome editing.
  • This technology holds promise for basic research and therapeutic applications in gene editing.