9T56

Cryo-EM structure of ISCro4-DBL-TBL-tDNA-dDNA synaptic complex

Summary for 9T56

• Entry DOI: 10.2210/pdb9t56/pdb
• EMDB information: 55572
• Descriptor: ISCro4 transposase, Target-binding loop (TBL) RNA, Donor-binding loop (DBL) RNA, ... (8 entities in total)
• Functional Keywords: nucleic acid interaction, rna binding, dna binding, tetramer, recombination, recombinase
• Biological source: Citrobacter rodentium ICC168; More
• Total number of polymer chains: 10
• Total formula weight: 264024.03

Authors

Fernandez Carrera, J.,Pelea, O.,Gerecke, S.E.,Chanez, C.,Jinek, M. (deposition date: 2025-11-04, release date: 2026-02-18)

Primary citation

Pelea, O.,Talas, A.,Carrera, J.F.,Mathis, N.,van de Venn, L.,Yeh, C.D.,Kulcsar, P.I.,Marquart, K.F.,Weber, Y.,Gerecke, S.E.,Harvey-Seutcheu, I.F.,Mailander, D.,Pfleiderer, M.M.,Chanez, C.,Corn, J.E.,Schwank, G.,Jinek, M.
Programmable genome editing in human cells using RNA-guided bridge recombinases.
Science, :eadz1884-eadz1884, 2026

PubMed Abstract: Site-specific insertion of gene-sized DNA fragments remains an unmet need in the genome editing field. IS110-family serine recombinases have recently been shown to mediate programmable DNA recombination in bacteria using a bispecific RNA guide (bridge RNA) that simultaneously recognizes target and donor sites. Here, we show that the bridge recombinase ISCro4 is highly active in human cells, and provide structural insights into its enhanced activity. Using plasmid- or all-RNA-based delivery, ISCro4 supports programmable multi-kilobase exisions and inversions, and facilitates donor DNA insertion at genomic sites with efficiencies exceeding 6%. Finally, we assess ISCro4 specificity and off-target activity. These results establish a framework for the development of bridge recombinases as next-generation tools for editing modalities that are beyond the capabilities of current technologies.

PubMed: 41642947
DOI: 10.1126/science.adz1884

Experimental method

ELECTRON MICROSCOPY (2.81 Å)