9nvu: Difference between revisions
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==Engineered OrufIscB-omegaRNA-target DNA complex== | |||
<StructureSection load='9nvu' size='340' side='right'caption='[[9nvu]], [[Resolution|resolution]] 2.71Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[9nvu]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Metagenome Metagenome] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=9NVU OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=9NVU 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]] 2.71Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=9nvu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=9nvu OCA], [https://pdbe.org/9nvu PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=9nvu RCSB], [https://www.ebi.ac.uk/pdbsum/9nvu PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=9nvu ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Naturally existing enzymes have been adapted for a variety of molecular technologies, with enhancements or modifications to the enzymes introduced to improve the desired function; however, it is difficult to engineer variants with enhanced activity while maintaining specificity. Here we engineer the compact Obligate Mobile Element Guided Activity (OMEGA) RNA-guided endonuclease IscB and its guiding RNA (omegaRNA) by combining ortholog screening, structure-guided protein domain design and RNA engineering, and deep learning-based structure prediction to generate an improved variant, NovaIscB. We show that the compact NovaIscB achieves up to 40% indel activity (~100-fold improvement over wild-type OgeuIscB) on the human genome with improved specificity relative to existing IscBs. We further show that NovaIscB can be fused with a methyltransferase to create a programmable transcriptional repressor, OMEGAoff, that is compact enough to be packaged in a single adeno-associated virus vector for persistent in vivo gene repression. This study highlights the power of combining natural diversity with protein engineering to design enhanced enzymes for molecular biology applications. | |||
Evolution-guided protein design of IscB for persistent epigenome editing in vivo.,Kannan S, Altae-Tran H, Zhu S, Xu P, Strebinger D, Oshiro R, Faure G, Moeller L, Pham J, Mears KS, Ni HM, Macrae RK, Zhang F Nat Biotechnol. 2025 May 7. doi: 10.1038/s41587-025-02655-3. PMID:40335752<ref>PMID:40335752</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 9nvu" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Metagenome]] | |||
[[Category: Synthetic construct]] | |||
[[Category: Xu P]] | |||
[[Category: Zhang F]] | |||
[[Category: Zhu S]] | |||
Latest revision as of 07:14, 3 June 2026
Engineered OrufIscB-omegaRNA-target DNA complex
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