Unique SiO2 Nanourchins Enable Amplification in Living Cells for In Situ Imaging of mRNAs

作者：Shen, Xiaotong; Wang, Yan; Zhang, Ying; Ouyang, Jin; Na, Na^*

来源：Advanced Functional Materials, 2018, 28(35): 1803286.

DOI：10.1002/adfm.201803286

• in situ imaging
• living cells
• messenger RNAs (mRNAs)
• reverse-transcription helicase-dependent isothermal amplification (RT-HDA)
• silica nanourchins

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.

• 出版日期2018-8-29
• 单位北京师范大学