View source for 9auf


==Cas9d 20bp R-loop Complex==
<StructureSection load='9auf' size='340' side='right'caption='[[9auf]], [[Resolution|resolution]] 2.73&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[9auf]] is a 5 chain structure with sequence from [https://en.wikipedia.org/wiki/Deltaproteobacteria Deltaproteobacteria]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=9AUF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=9AUF FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 2.73&#8491;</td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=9auf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=9auf OCA], [https://pdbe.org/9auf PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=9auf RCSB], [https://www.ebi.ac.uk/pdbsum/9auf PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=9auf ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/A0A1F8ZSN4_9DELT A0A1F8ZSN4_9DELT] 
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Type II CRISPR endonucleases are widely used programmable genome editing tools. Recently, CRISPR-Cas systems with highly compact nucleases have been discovered, including Cas9d (a type II-D nuclease). Here, we report the cryo-EM structures of a Cas9d nuclease (747 amino acids in length) in multiple functional states, revealing a stepwise process of DNA targeting involving a conformational switch in a REC2 domain insertion. Our structures provide insights into the intricately folded guide RNA which acts as a structural scaffold to anchor small, flexible protein domains for DNA recognition. The sgRNA can be truncated by up to ~25% yet still retain activity in vivo. Using ancestral sequence reconstruction, we generated compact nucleases capable of efficient genome editing in mammalian cells. Collectively, our results provide mechanistic insights into the evolution and DNA targeting of diverse type II CRISPR-Cas systems, providing a blueprint for future re-engineering of minimal RNA-guided DNA endonucleases.

DNA targeting by compact Cas9d and its resurrected ancestor.,Ocampo RF, Bravo JPK, Dangerfield TL, Nocedal I, Jirde SA, Alexander LM, Thomas NC, Das A, Nielson S, Johnson KA, Brown CT, Butterfield CN, Goltsman DSA, Taylor DW Nat Commun. 2025 Jan 7;16(1):457. doi: 10.1038/s41467-024-55573-4. PMID:39774105<ref>PMID:39774105</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 9auf" style="background-color:#fffaf0;"></div>
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Deltaproteobacteria]]
[[Category: Large Structures]]
[[Category: Bravo JPK]]
[[Category: Fregoso Ocampo R]]
[[Category: Taylor DW]]