中科院理化技术研究所科研成果 |新型RNA染料-CP——高选择性、低毒性-百灵威

中科院理化技术研究所科研成果 |新型RNA染料-CP——高选择性、低毒性

CP染料是一种新型RNA染料(发射波长为658nm,深红色荧光),可选择性地染色活细胞核仁中的RNA。由于在分子结构中引入了刚性共轭香豆素部分,使其显示出较大的Stokes位移(图1) [1]

图1 Fluorescent images of living cells by CP. (A) CP(5 μM) was incubated with Hela cells in DMEM for 20 min and imaged<sup> [1] </sup>.
图1 Fluorescent images of living cells by CP. (A) CP(5 μM) was incubated with Hela cells in DMEM for 20 min and imaged [1] .

细胞成像实验表明,CP染料可通过与核仁RNA结合,对活细胞核仁进行快速染色,性能优于市售的核仁染料SYTO RNA Select,具有较高的光稳定性和选择性(图2)。

图2 Cytotoxicity and photostability of CP. (a)Cytotoxicity of CP; (b)photostability of CP (5 μM) and SYTO RNASelect(1 μM) with increasing number of scans during cell imaging<sup> [1] </sup>.
图2 Cytotoxicity and photostability of CP. (a)Cytotoxicity of CP; (b)photostability of CP (5 μM) and SYTO RNASelect(1 μM) with increasing number of scans during cell imaging [1] .

由于上述优势及较低的细胞毒性(图2),CP染料能够监测有丝分裂过程中核仁RNA的变化(图3),使其成为实时、长期可视化核仁RNA动态变化的潜在工具。

图3 Dynamics ofnucleoliofstainedHeLacellsby CP during mitosis.(A–I) Time-dependent fluorescence imaging after addition of 5 mM CP<sup> [1] </sup>.
图3 Dynamics ofnucleoliofstainedHeLacellsby CP during mitosis.(A–I) Time-dependent fluorescence imaging after addition of 5 mM CP [1] .

顺铂是一种明星抗癌药物,研究人员用CP染料监测药物诱导的核仁RNA变化,如图4所示,监测5h后,对照组无明显变化,而实验组(顺铂处理的HeLa细胞中),由于RNA聚合酶I的重新分布,核仁在5小时后被包装成颗粒,核仁量减少,这与传统的银染色法结果非常吻合。同理,CP染料还可以监测其他抗癌药如放线菌素D和α-鹅膏蕈碱的抗癌效果,是实时、长期可视化核仁RNA动态变化的有力工具。

图4 Time-lapse imaging of nucleolus RNA in HeLa cells with cisplatin treated using CP (5 μM) in the range of hours<sup> [1] </sup>.
图4 Time-lapse imaging of nucleolus RNA in HeLa cells with cisplatin treated using CP (5 μM) in the range of hours [1] .
相关产品
CP RNA Dye, 98%
CP RNA染料
参考文献
  1. Zhou B, Liu W, Zhang H, et al. Imaging of nucleolar RNA in living cells using a highly photostable deep-red fluorescent probe[J]. Biosensors & Bioelectronics, 2015, 68:189-196.
相关阅读
中科院理化技术研究所科研成果 | 高选择性,高稳定性RNA染料—CP
低电渗合成琼脂糖——条带分离度提高近30%
CelRed、CelGreen核酸染料——安全无毒,不致癌
常用生化试剂一览
生化常用培养基及相关试剂的配制
常用生化试剂配制