Bright and Light-Up Sensing of Benzo[c,d]indole-oxazolopyridine Cyanine Dye for RNA and Its Application to Highly Sensitive Imaging of Nucleolar RNA in Living Cells.
Bright and Light-Up Sensing of Benzo[c,d]indole-oxazolopyridine Cyanine Dye for RNA and Its Application to Highly Sensitive Imaging of Nucleolar RNA in Living Cells.
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苯并[c,d]吲哚 - 恶唑并吡啶菁染料对RNA的明亮发光传感及其在活细胞中核仁RNA高灵敏度成像的应用
DOI：
10.1021/acsomega.2c02408
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发表时间：
2022-07-12
期刊：
影响因子：
4.1
通讯作者：
Nishizawa, Seiichi
中科院分区：
文献类型：
--
作者：
Higuchi, Kei;Sato, Yusuke;Togashi, Nao;Suzuki, Michiyuki;Yoshino, Yukina;Nishizawa, Seiichi
能够在与RNA结合时显示荧光信号响应的小分子探针在活细胞RNA成像方面很有前景。到目前为止，能够实现较大的荧光增强能力（>100倍）以及对RNA有较高的结合态量子产率（>0.50）的活细胞RNA成像探针鲜有报道。在此，苯并[c,d]吲哚 - 恶唑并吡啶（BIOP），一种不对称的单甲川菁类似物，被新开发为一种用于活细胞核仁RNA成像的明亮且具有较大荧光增强效果的探针。BIOP作为一种黄色发射探针（发射波长λem = 570 nm），在与RNA结合时表现出显著的荧光增强响应（770倍），且结合态量子产率较高（0.52）。通过对RNA荧光特性的系统比较，我们证明了BIOP相对于市售的RNA染色探针SYTO RNA select在稳健和灵敏的RNA检测方面的优势。此外，发现BIOP在活细胞中具有高膜通透性和低细胞毒性。活细胞成像检测表明，BIOP在与核仁RNA结合时在核仁中显示出的发射比SYTO RNA select强得多。此外，BIOP有助于对核仁RNA进行高灵敏度成像，其中50 nM的BIOP在孵育20分钟后即可对活细胞中的核仁RNA进行染色。
Small molecular weight probes that can show a fluorescence signaling response upon binding to RNAs are promising for RNA imaging in living cells. Live-cell RNA imaging probes that can achieve a large light-up ability (>100-fold) and high Φbound value for RNA (>0.50) have been rarely reported to date. Here, benzo[c,d]indole-oxazolopyridine (BIOP), an unsymmetrical monomethine cyanine analogue, was newly developed as a bright and large light-up probe for imaging of nucleolar RNA in living cells. BIOP served as a yellow-emissive probe (λem = 570 nm) and exhibited a significant light-up response upon RNA binding (770-fold) with a high Φbound value (0.52). We demonstrated the advantages of BIOP over a commercially available RNA-staining probe, SYTO RNA select, for robust and sensitive RNA sensing by a systematic comparison of fluorescent properties for RNA. In addition, BIOP was found to possess high membrane permeability and low cytotoxicity in living cells. The examination of live-cell imaging revealed that BIOP exhibited emission in the nucleolus upon binding to nucleolar RNA much stronger than that of SYTO RNA select. Furthermore, BIOP facilitated the highly sensitive imaging of nucleolar RNA, in which 50 nM BIOP can stain nucleolar RNA in living cells with a 20 min incubation.
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