1/1/2024 0 Comments Zenofae gene guideMcGovern Institute for Brain Research at MIT, Cambridge, MA, USAĭepartment of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA, USAĭepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA Makoto Saito, Peiyu Xu, Guilhem Faure, Samantha Maguire, Soumya Kannan, Han Altae-Tran, Sam Vo, AnAn Desimone, Rhiannon K. These authors contributed equally: Makoto Saito, Peiyu Xuīroad Institute of MIT and Harvard, Cambridge, MA, USA Our results show that Fz is a eukaryotic OMEGA system, demonstrating that RNA-guided endonucleases are present in all three domains of life. Finally, we resolved the structure of Spizellomyces punctatus Fz (SpuFz) at 2.7Å using cryogenic-electron microscopy, revealing the conservation of the core regions among Fz, TnpB and Cas12, despite diverse cognate RNA structures. We also show that Fz can be reprogrammed for human genome engineering applications. Here we report the biochemical characterization of Fz, showing that it is an RNA-guided DNA endonuclease. TnpB may also be the ancestor of the eukaryotic transposon-encoded Fanzor (Fz) proteins 4,7, raising the possibility that eukaryotes are also equipped with CRISPR-Cas/OMEGA-like programmable RNA-guided endonucleases. The OMEGA effector TnpB is the putative ancestor of Cas12 and has RNA-guided endonuclease activity 4,6. Recently, a new class of prokaryotic RNA-guided system (termed OMEGA) was reported 4,5. Although a few eukaryotic RNA-guided systems have been studied so far, such as RNA interference 2 and ribosomal RNA modification 3, it remains unclear whether eukaryotes have RNA-guided endonucleases. Cas effectors, like Cas9 and Cas12, perform guide RNA-dependent DNA cleavage 1. For example, the prokaryotic CRISPR-Cas systems provide adaptive immunity for bacteria and archaea against foreign genetic elements. RNA-guided systems, which employ complementarity between a guide RNA and target nucleic acid sequences for recognition of genetic elements, play a central role in biological processes in both prokaryotes and eukaryotes.
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