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Földes-Papp, Z; König, K; Studier, H; Bückle, R; Breunig, HG; Uchugonova, A; Kostner, GM.
Trafficking of mature miRNA-122 into the nucleus of live liver cells.
Curr Pharm Biotechnol. 2009; 10(6): 569-578. Doi: 10.2174/138920109789069332
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Führende Autor*innen der Med Uni Graz
Földes-Papp Zeno
Co-Autor*innen der Med Uni Graz
Kostner Gerhard
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Abstract:
The binding of superquencher molecular beacon (SQMB) probes to human single-stranded cellular miRNA-122 targets was detected in various single live cells with femtosecond laser microscopy. For delivery of the SQMB-probes, 3D-nanoprocessing of single cells with sub-15 femtosecond 85 MHz near-infrared laser pulses was applied. Transient nanopores were formed by focusing the laser beam for some milliseconds on the membrane of a single cell in order to import of SQMB-probes into the cells. In single cells of the human liver cell lines Huh-7D12 and IHH that expressed miRNA-122, we measured target binding in the cytoplasm by two-photon fluorescence imaging. We found increased fluorescence with time in a nonlinear manner up to the point where steady state saturation was reached. We also studied the intracellular distribution of target SQMB and provide for the first time strong experimental evidence that cytoplasmic miRNA travels into the cell nucleus. To interpret nonlinear binding, a number of individual miRNA-122 positive cells (Huh-7D12 and IHH) and negative control cells, human VA13 fibroblasts and Caco-2 cells were analyzed. Our experimental data are consistent with the cytoplasmic assembly of nuclear miRNA and provide further mechanistic insight in the regulatory function of miRNAs in cellular physiology. An open issue in the regulation of gene expression by miRNA is whether miRNA can activate gene expression in addition to the well-known inhibitory effect. A first step for such a regulatory role could be the travelling of miRNA-RISC into the nucleus.
Find related publications in this database (using NLM MeSH Indexing)
Biological Transport, Active - physiology
Cell Line -
Cell Nucleus - metabolism Cell Nucleus - ultrastructure
Hepatocytes - metabolism Hepatocytes - ultrastructure
Humans -
MicroRNAs - pharmacokinetics
Microscopy, Fluorescence, Multiphoton - methods

Find related publications in this database (Keywords)
Single live cells
cell trafficking
micro-RNA
RISC
nuclear micro-RNA-122
cytoplasmic assembly of nuclear miRNA-122
cell biology theory of miRNA role and function
activation of gene expression
in vivo two-photon imaging
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