8GKH

Structure of the Spizellomyces punctatus Fanzor (SpuFz) in complex with omega RNA and target DNA

Summary for 8GKH

• Entry DOI: 10.2210/pdb8gkh/pdb
• EMDB information: 40184
• Descriptor: DNA (5'-D(P*CP*GP*GP*TP*AP*CP*CP*CP*GP*GP*GP*CP*AP*TP*A)-3'), Maltodextrin-binding protein (Fragment),OrfB_Zn_ribbon domain-containing protein, DNA (35-MER), ... (7 entities in total)
• Functional Keywords: fanzor, eukaryotic, rna-guided, nuclease, gene editing, rna binding protein, rna binding protein-rna-dna complex, rna binding protein/rna/dna
• Biological source: Methanosarcina mazei; More
• Total number of polymer chains: 4
• Total formula weight: 158527.65

Authors

Xu, P.,Saito, M.,Zhang, F. (deposition date: 2023-03-19, release date: 2023-07-05, Last modification date: 2023-08-30)

Primary citation

Saito, M.,Xu, P.,Faure, G.,Maguire, S.,Kannan, S.,Altae-Tran, H.,Vo, S.,Desimone, A.,Macrae, R.K.,Zhang, F.
Fanzor is a eukaryotic programmable RNA-guided endonuclease.
Nature, 620:660-668, 2023

PubMed Abstract: 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.

PubMed: 37380027
DOI: 10.1038/s41586-023-06356-2

Experimental method

ELECTRON MICROSCOPY (2.7 Å)