8T2D

Ubiquitin variant i53:Mutant T12Y.T14E.L67R with 53BP1 Tudor domain

8T2D の概要

• エントリーDOI: 10.2210/pdb8t2d/pdb
• 関連するPDBエントリー: 8SVG 8SVH 8SVI 8SVJ
• 分子名称: Ubiquitin variant i53, Tumor protein p53 binding protein 1 (3 entities in total)
• 機能のキーワード: inhibitor, protein binding
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 22710.78

構造登録者

Partridge, J.R.,Holden, J.K.,Wibowo, A.S.,Mulichak, A. (登録日: 2023-06-05, 公開日: 2024-03-27, 最終更新日: 2024-04-03)

主引用文献

Perez-Bermejo, J.A.,Efagene, O.,Matern, W.M.,Holden, J.K.,Kabir, S.,Chew, G.M.,Andreoletti, G.,Catton, E.,Ennis, C.L.,Garcia, A.,Gerstenberg, T.L.,Hill, K.A.,Jain, A.,Krassovsky, K.,Lalisan, C.D.,Lord, D.,Quejarro, B.J.,Sales-Lee, J.,Shah, M.,Silva, B.J.,Skowronski, J.,Strukov, Y.G.,Thomas, J.,Veraz, M.,Vijay, T.,Wallace, K.A.,Yuan, Y.,Grogan, J.L.,Wienert, B.,Lahiri, P.,Treusch, S.,Dever, D.P.,Soros, V.B.,Partridge, J.R.,Seim, K.L.
Functional screening in human HSPCs identifies optimized protein-based enhancers of Homology Directed Repair.
Nat Commun, 15:2625-2625, 2024

PubMed Abstract: Homology Directed Repair (HDR) enables precise genome editing, but the implementation of HDR-based therapies is hindered by limited efficiency in comparison to methods that exploit alternative DNA repair routes, such as Non-Homologous End Joining (NHEJ). In this study, we develop a functional, pooled screening platform to identify protein-based reagents that improve HDR in human hematopoietic stem and progenitor cells (HSPCs). We leverage this screening platform to explore sequence diversity at the binding interface of the NHEJ inhibitor i53 and its target, 53BP1, identifying optimized variants that enable new intermolecular bonds and robustly increase HDR. We show that these variants specifically reduce insertion-deletion outcomes without increasing off-target editing, synergize with a DNAPK inhibitor molecule, and can be applied at manufacturing scale to increase the fraction of cells bearing repaired alleles. This screening platform can enable the discovery of future gene editing reagents that improve HDR outcomes.

PubMed: 38521763
DOI: 10.1038/s41467-024-46816-5

実験手法

X-RAY DIFFRACTION (1.751 Å)