دورية أكاديمية
Multiplex, Quantitative, High-Resolution Imaging of Protein:Protein Complexes via Hybridization Chain Reaction
| العنوان: | Multiplex, Quantitative, High-Resolution Imaging of Protein:Protein Complexes via Hybridization Chain Reaction |
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| المؤلفون: | Samuel J. Schulte, Boyoung Shin, Ellen V. Rothenberg, Niles A. Pierce |
| سنة النشر: | 2024 |
| مصطلحات موضوعية: | Biophysics, Biochemistry, Medicine, Cell Biology, Genetics, Molecular Biology, Physiology, Biotechnology, Immunology, Developmental Biology, Cancer, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, Physical Sciences not elsewhere classified, two target proteins, target classes simultaneously, reagents bind nonspecifically, precise relative quantitation, hybridization chain reaction, highly autofluorescent formalin, gene regulatory networks, facilitates spatial exploration, single amplified signal, mouse prot cells, dependent cooperative probes, three different protein, hcr enables accurate, generate amplified background, protein complexes simultaneously, demonstrate multiplex imaging |
| الوصف: | Signal amplification based on the mechanism of hybridization chain reaction (HCR) facilitates spatial exploration of gene regulatory networks by enabling multiplex, quantitative, high-resolution imaging of RNA and protein targets. Here, we extend these capabilities to the imaging of protein:protein complexes, using proximity-dependent cooperative probes to conditionally generate a single amplified signal if and only if two target proteins are colocalized within the sample. HCR probes and amplifiers combine to provide automatic background suppression throughout the protocol, ensuring that even if reagents bind nonspecifically in the sample, they will not generate amplified background. We demonstrate protein:protein imaging with a high signal-to-background ratio in human cells, mouse proT cells, and highly autofluorescent formalin-fixed paraffin-embedded (FFPE) human breast tissue sections. Further, we demonstrate multiplex imaging of three different protein:protein complexes simultaneously and validate that HCR enables accurate and precise relative quantitation of protein:protein complexes with subcellular resolution in an anatomical context. Moreover, we establish a unified framework for simultaneous multiplex, quantitative, high-resolution imaging of RNA, protein, and protein:protein targets, with one-step, isothermal, enzyme-free HCR signal amplification performed for all target classes simultaneously. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| العلاقة: | https://figshare.com/articles/journal_contribution/Multiplex_Quantitative_High-Resolution_Imaging_of_Protein_Protein_Complexes_via_Hybridization_Chain_Reaction/25016288Test |
| DOI: | 10.1021/acschembio.3c00431.s001 |
| الإتاحة: | https://doi.org/10.1021/acschembio.3c00431.s001Test https://figshare.com/articles/journal_contribution/Multiplex_Quantitative_High-Resolution_Imaging_of_Protein_Protein_Complexes_via_Hybridization_Chain_Reaction/25016288Test |
| حقوق: | CC BY-NC 4.0 |
| رقم الانضمام: | edsbas.7702E78 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1021/acschembio.3c00431.s001 |
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