Fanzor is a eukaryotic programmable RNA-guided endonuclease
Makoto Saito1,2,3,4,5, Peiyu Xu1,2,3,4,5, Guilhem Faure1,2,3,4,5
1Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Nature
|June 28, 2023
Summary
Eukaryotes possess RNA-guided endonucleases, exemplified by Fanzor (Fz) proteins, which are crucial for genome engineering. This discovery confirms RNA-guided systems across all three domains of life.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- RNA-guided systems, like CRISPR-Cas, are vital in prokaryotes for adaptive immunity and DNA cleavage.
- Eukaryotic RNA-guided systems are less understood, with no confirmed RNA-guided endonucleases.
- Prokaryotic OMEGA systems, including TnpB, share ancestry with eukaryotic Fanzor (Fz) proteins.
Purpose of the Study:
- To biochemically characterize Fanzor (Fz) proteins.
- To investigate Fz's potential for reprogramming in human genome engineering.
- To elucidate the structural basis of Fz's RNA-guided DNA endonuclease activity.
Main Methods:
- Biochemical assays to determine Fz's nuclease activity.
- Reprogramming Fz for targeted DNA cleavage in human cells.
- Cryogenic electron microscopy (cryo-EM) for structural analysis of Fz.
Main Results:
- Fanzor (Fz) proteins function as RNA-guided DNA endonucleases.
- Fz can be successfully engineered for genome editing applications.
- Structural analysis reveals conserved regions between Fz, TnpB, and Cas12.
Conclusions:
- Fanzor (Fz) represents a eukaryotic OMEGA system.
- RNA-guided endonucleases are present in all three domains of life (bacteria, archaea, and eukaryotes).
- Fz expands the toolkit for eukaryotic genome engineering.
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