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Symmetry-breaking malachite green as a near-infrared light-activated fluorogenic photosensitizer for RNA proximity labeling.

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  • معلومة اضافية
    • المصدر:
      Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE
    • بيانات النشر:
      Publication: 1992- : Oxford : Oxford University Press
      Original Publication: London, Information Retrieval ltd.
    • الموضوع:
    • نبذة مختصرة :
      Cellular RNA is asymmetrically distributed in cells and the regulation of RNA localization is crucial for proper cellular functions. However, limited chemical tools are available to capture dynamic RNA localization in complex biological systems with high spatiotemporal resolution. Here, we developed a new method for RNA proximity labeling activated by near-infrared (NIR) light, which holds the potential for deep penetration. Our method, termed FAP-seq, utilizes a genetically encoded fluorogen activating protein (FAP) that selectively binds to a set of substrates known as malachite green (MG). FAP binding restricts the rotation of MG and rapidly activates its fluorescence in a wash-free manner. By introducing a monoiodo modification to MG, we created a photosensitizer (MG-HI) with the highest singlet oxygen generation ability among various MG derivatives, enabling both protein and RNA proximity labeling in live cells. New insights are provided in the transcriptome analysis with FAP-seq, while a deeper understanding of the symmetry-breaking structural arrangement of FAP-MG-HI was obtained through molecular dynamics simulations. Overall, our wash-free and NIR light-inducible RNA proximity labeling method (FAP-seq) offers a powerful and versatile approach for investigating complex mechanisms underlying RNA-related biological processes.
      (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)
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    • Grant Information:
      R35 GM133385 United States GM NIGMS NIH HHS; R35GM13385 United States NH NIH HHS
    • الرقم المعرف:
      12058M7ORO (malachite green)
      0 (Rosaniline Dyes)
      0 (Photosensitizing Agents)
      0 (Fluorescent Dyes)
      63231-63-0 (RNA)
      17778-80-2 (Singlet Oxygen)
    • الموضوع:
      Date Created: 20240226 Date Completed: 20240424 Latest Revision: 20241018
    • الموضوع:
      20241018
    • الرقم المعرف:
      PMC11040151
    • الرقم المعرف:
      10.1093/nar/gkae125
    • الرقم المعرف:
      38407347