首页> 外文期刊>Analytical chemistry >Biomineralized Metal-Organic Framework Nanoparticles Enable Enzymatic Rolling Circle Amplification in Living Cells for Ultrasensitive MicroRNA Imaging
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Biomineralized Metal-Organic Framework Nanoparticles Enable Enzymatic Rolling Circle Amplification in Living Cells for Ultrasensitive MicroRNA Imaging

机译:生物丙原化金属 - 有机框架纳米粒子使活细胞酶轧制圆形扩增能够进行超细微征微瘤成像

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摘要

The enzymatic amplification strategy in living cells faces challenges of highly efficient intracellular codelivery of amplification reagents including DNA polymerase. In this work, we develop biomineralized metal-organic framework nanoparticles (MOF NPs) as a carrier system for intracellular codelivery of phi 29 DNA polymerase (phi 29DP) and nucleic acid probes and realize a polymerization amplification reaction in living cells. A pH-sensitive biodegradable MOF NP of zeolitic imidazolate framework-8 (ZIF-8) is utilized to encapsulate phi 29DP and adsorb nucleic acid probes. After uptake into cells, the encapsulated phi 29DP and surface-adsorbed DNA probes are released and escaped from endolysosomes. In the presence of phi 29DP and deoxyribonucleotide triphosphates (dNTPs), the intracellular miRNA-21 triggers a rolling circle amplification (RCA) reaction and the autonomous synthesized Mg2+-dependent DNAzyme cleaves the fluorogenic substrate, providing a readout fluorescence signal for the monitoring of miRNA-21. This is the first example of the intracellular RCA reaction in living cells. Therefore, the proposed method provides new opportunities for achieving enzymatic amplification reaction in living cells.
机译:活细胞中的酶促扩增策略面临高效细胞内编码的挑战,扩增试剂包括DNA聚合酶。在这项工作中,我们将生物丙原化金属 - 有机骨架纳米颗粒(MOF NPS)作为载体系统作为载体系统,用于PHI 29 DNA聚合酶(PHI 29DP)和核酸探针,并在活细胞中实现聚合扩增反应。利用沸石咪唑酯框架-8(ZIF-8)的pH敏感的可生物降解的MOF NP(ZIF-8)来包封PHI 29DP和吸附核酸探针。在进入细胞后,封装的PHI 29DP和表面吸附的DNA探针被释放并从底糖体中逸出。在PHI 29DP和脱氧核糖核苷酸的三磷酸(DNTPS)存在下,细胞内miRNA-21触发滚动圆扩增(RCA)反应,并且自主合成的Mg2 +彼义的DNazyme切割荧光底物,为miRNA的监测提供读出荧光信号。 -21。这是活细胞中细胞内RCA反应的第一个例子。因此,所提出的方法提供了实现活细胞中酶促扩增反应的新机会。

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  • 来源
    《Analytical chemistry》 |2019年第14期|共9页
  • 作者单位

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

    Hunan Univ Coll Chem &

    Chem Engn Key Lab Chemo Biosensing &

    Chemometr Changsha 410082 Hunan Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

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