ArticleNucleic acids research2021
Molecular underpinnings of ssDNA specificity by Rep HUH-endonucleases and implications for HUH-tag multiplexing and engineering.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 51 citations in OpenAlex.
- Homogeneous Antibody-DNA Conjugates Using Unmodified Oligonucleotides and Photo-Cross-Linkable Protein G-HUH Endonuclease Fusion Proteins.Bioconjugate chemistry · 2026Article
- Sequence-directed covalent protein-RNA linkages in a single step using engineered HUH-tags.Nucleic acids research · 2026Article
- HUH-tagged Cas9 as a platform for efficient ssODN-mediated knock-in via embryo and adult injection in insects.Communications biology · 2026Article
- Structures of nucleotide-bound Redondovirus Rep protein link conformation and function.PLoS pathogens · 2026Article
- Reversible Sandwich-Based Particle Nanoswitch for Continuous Protein Monitoring at Picomolar Concentrations with Automated Calibration.Angewandte Chemie (International ed. in English) · 2025Article
- Identification and biochemical characterization of a novel porcine circovirus associated with porcine respiratory and diarrheal diseases.Microbiology spectrum · 2025Article
- A streamlined base editor engineering strategy to reduce bystander editing.Nature communications · 2025Article
- High-throughput Flow-cytometry Measurement of Cellular Mechanotype Based on Rupture and Delivery of DNA Tension Probes into Cells.Journal of visualized experiments : JoVE · 2025Article
- Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.Nature biotechnology · 2025Article
- Cas9-Rep fusion tethers donor DNA in vivo and boosts the efficiency of HDR-mediated genome editing.Plant biotechnology journal · 2025Article
- Directed evolution of a sequence-specific covalent protein tag for RNA labeling.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- The replication protein of duck circovirus unwinds dsDNA in an ATP-driven and metal ion-dependent manner from 3' to 5'.Frontiers in veterinary science · 2025Article
- HUHgle: An Interactive Substrate Design Tool for Covalent Protein-ssDNA Labeling Using HUH-Tags.ACS synthetic biology · 2024Article
- Review
- Computational studies on rep and capsid proteins of CRESS DNA viruses.Virusdisease · 2024Article
- Exploration and Application of DNA-Binding Proteins to Make a Versatile DNA-Protein Covalent-Linking Patch (D-Pclip): The Case of a Biosensing Element.Journal of the American Chemical Society · 2024Article
- Multiparametric domain insertional profiling of adeno-associated virus VP1.Molecular therapy. Methods & clinical development · 2023Article
- Article
- The crystal structure of the human smacovirus 1 Rep domain.Acta crystallographica. Section F, Structural biology communications · 2023Article
- Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.bioRxiv : the preprint server for biology · 2023Article
Corrections and comments
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Authors and funding
13 authors at 1 institution in 1 country.
Funding
Abstract
Replication initiator proteins (Reps) from the HUH-endonuclease superfamily process specific single-stranded DNA (ssDNA) sequences to initiate rolling circle/hairpin replication in viruses, such as crop ravaging geminiviruses and human disease causing parvoviruses. In biotechnology contexts, Reps are the basis for HUH-tag bioconjugation and a critical adeno-associated virus genome integration tool. We solved the first co-crystal structures of Reps complexed to ssDNA, revealing a key motif for conferring sequence specificity and for anchoring a bent DNA architecture. In combination, we developed a deep sequencing cleavage assay, termed HUH-seq, to interrogate subtleties in Rep specificity and demonstrate how differences can be exploited for multiplexed HUH-tagging. Together, our insights allowed engineering of only four amino acids in a Rep chimera to predictably alter sequence specificity. These results have important implications for modulating viral infections, developing Rep-based genomic integration tools, and enabling massively parallel HUH-tag barcoding and bioconjugation applications.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.