Unique SiO2 Nanourchins Enable Amplification in Living Cells for In Situ Imaging of mRNAs,Advanced Functional Materials

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Unique SiO2 Nanourchins Enable Amplification in Living Cells for In Situ Imaging of mRNAs
Advanced Functional Materials ( IF 18.5 ) Pub Date : 2018-07-12 , DOI: 10.1002/adfm.201803286
Xiaotong Shen ₁ , Yan Wang ₁ , Ying Zhang ₁ , Jin Ouyang ₁ , Na Na ₁

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Sufficient delivery of nucleic acid reagents into cells for amplifications with great activities would be significant for in situ intracellular imaging of messenger RNAs in living cells, showing great potentials in chemistry and biology. Herein, SiO₂ nanourchins (SNUs) are developed, with two‐pronged pair of primers anchored on enzyme‐functionalized mesoporous SiO₂ cores, for in situ imaging of mRNAs in living cells. Endowed by the urchins‐like structure to represent high loading capacity, efficient activity without mutual interference, and sufficient delivery into cells, SNUs maintain sustainable reactivities for amplification through primers' 3′‐end complete exposure and synergistic reagents release by increased dissociation or mobility during hybridization. “One‐step” reverse‐transcription helicase‐dependent isothermal amplifications are employed in living cells with Klenow (exo⁻) DNA polymerase to accomplish reverse transcriptions and polymerization amplifications. With high efficiency and stability, low toxicity, and good specificity, this method can detect amol of mRNA in dozens of microliter samples (<0.02 molecules per cell) in living cells, providing an ultrasensitive tool for fundamental understanding of mRNAs in cellular processes.

中文翻译：

独特的SiO2纳米urchins可在活细胞中扩增mRNA的原位成像

对于活细胞中信使RNA的原位细胞内成像，将核酸试剂充分递送到细胞中以进行具有很大活性的扩增将具有重要意义，在化学和生物学领域显示出巨大的潜力。本文中，开发了SiO ₂纳米顽童（SNU），其中两对引物锚定在酶功能化介孔SiO _2上核心，用于活细胞中mRNA的原位成像。SNU具有类似海胆的结构，可表现出高负载能力，有效的活性而不会相互干扰，并且能充分递送到细胞中，可通过引物的3'端完全暴露和协同试剂的释放来维持可持续的反应性，并通过增加解离或迁移来释放协同试剂。杂交。“一步法”逆转录解旋酶依赖的等温扩增是在活细胞用Klenow（外使用^-DNA聚合酶可完成逆转录和聚合扩增。该方法具有高效率和稳定性，低毒性和良好的特异性，可检测活细胞中数十微升样品（每个细胞<0.02分子）中的amol mRNA，为基础了解细胞过程中的mRNA提供了超灵敏的工具。

更新日期：2018-07-12

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