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TitleFanzor is a eukaryotic programmable RNA-guided endonuclease.
Journal, issue, pagesNature, Vol. 620, Issue 7974, Page 660-668, Year 2023
Publish dateJun 28, 2023
AuthorsMakoto Saito / Peiyu Xu / Guilhem Faure / Samantha Maguire / Soumya Kannan / Han Altae-Tran / Sam Vo / AnAn Desimone / Rhiannon K Macrae / Feng Zhang /
PubMed AbstractRNA-guided systems, which use complementarity between a guide RNA and target nucleic acid sequences for recognition of genetic elements, have a central role in biological processes in both ...RNA-guided systems, which use complementarity between a guide RNA and target nucleic acid sequences for recognition of genetic elements, have a central role in biological processes in both prokaryotes and eukaryotes. For example, the prokaryotic CRISPR-Cas systems provide adaptive immunity for bacteria and archaea against foreign genetic elements. Cas effectors such as Cas9 and Cas12 perform guide-RNA-dependent DNA cleavage. Although a few eukaryotic RNA-guided systems have been studied, including RNA interference and ribosomal RNA modification, it remains unclear whether eukaryotes have RNA-guided endonucleases. Recently, a new class of prokaryotic RNA-guided systems (termed OMEGA) was reported. The OMEGA effector TnpB is the putative ancestor of Cas12 and has RNA-guided endonuclease activity. TnpB may also be the ancestor of the eukaryotic transposon-encoded Fanzor (Fz) proteins, raising the possibility that eukaryotes are also equipped with CRISPR-Cas or OMEGA-like programmable RNA-guided endonucleases. Here we report the biochemical characterization of Fz, showing that it is an RNA-guided DNA endonuclease. We also show that Fz can be reprogrammed for human genome engineering applications. Finally, we resolve the structure of Spizellomyces punctatus Fz at 2.7 Å using cryogenic electron microscopy, showing the conservation of core regions among Fz, TnpB and Cas12, despite diverse cognate RNA structures. Our results show that Fz is a eukaryotic OMEGA system, demonstrating that RNA-guided endonucleases are present in all three domains of life.
External linksNature / PubMed:37380027 / PubMed Central
MethodsEM (single particle)
Resolution2.7 Å
Structure data

40184

8gkh

EMDB-40184, PDB-8gkh:
Structure of the Spizellomyces punctatus Fanzor (SpuFz) in complex with omega RNA and target DNA
Method: EM (single particle) / Resolution: 2.7 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-HOH:
WATER / Water

Source
  • spizellomyces punctatus (fungus)
  • synthetic construct (others)
  • methanosarcina mazei (archaea)
KeywordsRNA BINDING PROTEIN/RNA/DNA / Fanzor / Eukaryotic / RNA-guided / nuclease / Gene editing / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA-DNA complex

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