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Unique SiO 2 Nanourchins Enable Amplification in Living Cells for In Situ Imaging of mRNAs
Author(s) -
Shen Xiaotong,
Wang Yan,
Zhang Ying,
Ouyang Jin
Publication year - 2018
Publication title -
advanced functional materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.069
H-Index - 322
eISSN - 1616-3028
pISSN - 1616-301X
DOI - 10.1002/adfm.201803286
Subject(s) - loop mediated isothermal amplification , in situ , exonuclease , nuclease , biophysics , reverse transcriptase , microbiology and biotechnology , dna , biology , polymerase , materials science , nanotechnology , rna , chemistry , biochemistry , organic chemistry , gene
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, SiO 2 nanourchins (SNUs) are developed, with two‐pronged pair of primers anchored on enzyme‐functionalized mesoporous SiO 2 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.