ArticleBiochemistry and biophysics reports2017
Application of a novel HiBiT peptide tag for monitoring ATF4 protein expression in Neuro2a cells.
Article in Biochemistry and biophysics reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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37 citing papers in PubMed, 53 citations in OpenAlex.
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- Optimization and Characterization of SHIP1 Ligands for Cellular Target Engagement and Activity in Alzheimer's Disease Models.Journal of medicinal chemistry · 2026Article
- Binding of Glycyl-tRNA synthetase to Mengovirus RNA stimulates translation.Nucleic acids research · 2026Article
- Optimization and Characterization of SHIP1 Ligands for Cellular Target Engagement and Activity in Alzheimer's Disease Models.bioRxiv : the preprint server for biology · 2026Article
- A trove of antiviral TRIM family E3 ligases in reptiles.bioRxiv : the preprint server for biology · 2025Article
- A flexible, high-throughput system for studying live mRNA translation with HiBiT technology.Nucleic acids research · 2025Article
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- Poly-GR repeats associated with ALS/FTD geneScience signaling · 2024Article
- Virus-derived circular RNAs populate hepatitis C virus-infected cells.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- CRISPR-Cas9-Mediated Bioluminescent Tagging of Endogenous Proteins by Fluorescent Protein-Assisted Cell Sorting.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Four-color single-molecule imaging system for tracking GPCR dynamics with fluorescent HiBiT peptide.Biophysics and physicobiology · 2024Article
- A New Derivative of Retro-2 Displays Antiviral Activity against Respiratory Syncytial Virus.International journal of molecular sciences · 2023Article
- Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation ofiScience · 2023Article
- Prediction of prime editing insertion efficiencies using sequence features and DNA repair determinants.Nature biotechnology · 2023Article
- Specific, sensitive and quantitative protein detection by in-gel fluorescence.Nature communications · 2023Article
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- Prime editing for precise and highly versatile genome manipulation.Nature reviews. Genetics · 2023Review
- Investigation of adenosine A1 receptor-mediated β-arrestin 2 recruitment using a split-luciferase assay.Frontiers in pharmacology · 2023Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A split NanoLuc assay system consisting of two fragments, large N-terminal and small C-terminal regions (NanoBiT), was developed to investigate protein-protein interactions within living cells. Interestingly, the replacement of five amino acids among 11 C-terminal amino acids dramatically increased affinity against the large N-terminal fragment, LgBiT, and the complex had NanoLuc luciferase activity. In this study, we first applied this small fragment, HiBiT, to elucidate the expression of ATF4 protein by transient overexpression of HiBiT-tagged ATF4. According to the regulation of intrinsic ATF4 protein, stabilization of HiBiT-tagged ATF4 with a proteasome inhibitor, MG132, was observed by detecting luciferase activity in cell lysate and after SDS-PAGE and transfer onto a PVDF membrane. Next, we knocked-in the HiBiT-epitope tag into the ATF4 gene using the CRISPR/Cas9 system and rapidly selected positive clones by measuring luciferase activity in an aliquot of each cell suspension. Using a selected clone, we observed that the expression of HiBiT-tagged ATF4 in the selected cells varied in response to treatment with protein synthesis inhibitors or proteasome inhibitors and tunicamycin. Altogether, this novel HiBiT tag is a useful tool to evaluate the endogenous expression levels of proteins of interest.
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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.