ArticleCancers2021
A Novel Nanobody Precisely Visualizes Phosphorylated Histone H2AX in Living Cancer Cells under Drug-Induced Replication Stress.
Article in Cancers, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 29 citations in OpenAlex.
- Post-translational modifications in the oral microenvironment: Stem cell regulation from periodontal regeneration to oral cancer therapy.World journal of stem cells · 2025Review
- Engineered chromatin readers track damaged chromatin dynamics in live cells and animals.Nature communications · 2025Article
- Inducible tag-free degradation of endogenous proteins with AlissAID and development of a photoactivable inducer.Communications biology · 2025Article
- NanoTag is an improved method to map interactions between DNA and proteins not requiring IgG.Scientific reports · 2025Article
- Neutralizing Nanobodies against Venoms fromToxins · 2024Article
- Uptake of substances into living mammalian cells by microwave induced perturbation of the plasma membrane.Scientific reports · 2024Article
- Characterization of novel CD19-specific VHHs isolated from a camelid immune library by phage display.Journal of translational medicine · 2023Article
- Wnt3a signaling with serum supply induces replication stress in cultured cells.Biochemistry and biophysics reports · 2023Article
- Emerging Landscape of Nanobodies and Their Neutralizing Applications against SARS-CoV-2 Virus.ACS pharmacology & translational science · 2023Review
- Modular Site-Specific Conjugation of Nanobodies Using Two Co-Associating Tags.International journal of molecular sciences · 2022Article
- Article
- Downstream events initiated by expression of FSHD-associated DUX4: Studies of nucleocytoplasmic transport, γH2AX accumulation, and Bax/Bak-dependence.Biology open · 2022Article
- Molecular probes for cellular imaging of post-translational proteoforms.RSC chemical biology · 2022Review
- Preparation of a Nanobody Specific to Dectin 1 and Its Anti-inflammatory Effects on Fungal Keratitis.International journal of nanomedicine · 2022Article
- Creating a Selective Nanobody Against 3-Nitrotyrosine Containing Proteins.Frontiers in chemistry · 2022Article
Corrections and comments
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Authors and funding
14 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Histone H2AX phosphorylated at serine 139 (γ-H2AX) is a hallmark of DNA damage, signaling the presence of DNA double-strand breaks and global replication stress in mammalian cells. While γ-H2AX can be visualized with antibodies in fixed cells, its detection in living cells was so far not possible. Here, we used immune libraries and phage display to isolate nanobodies that specifically bind to γ-H2AX. We solved the crystal structure of the most soluble nanobody in complex with the phosphopeptide corresponding to the C-terminus of γ-H2AX and show the atomic constituents behind its specificity. We engineered a bivalent version of this nanobody and show that bivalency is essential to quantitatively visualize γ-H2AX in fixed drug-treated cells. After labelling with a chemical fluorophore, we were able to detect γ-H2AX in a single-step assay with the same sensitivity as with validated antibodies. Moreover, we produced fluorescent nanobody-dTomato fusion proteins and applied a transduction strategy to visualize with precision γ-H2AX foci present in intact living cells following drug treatment. Together, this novel tool allows performing fast screenings of genotoxic drugs and enables to study the dynamics of this particular chromatin modification in individual cancer cells under a variety of conditions.
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