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The capability to profile small RNAs responsible for post-transcriptional regulation of genes expression is essential for molecular identification and characterization of cellular phenotypes. However, compared to long RNA, the quantification of small RNA, such as microRNAs (miRNA), in live cells is still challenging. A group led by Prof. SHI Peng develops an intracellular biopsy technique for fast, multiplexed and highly-sensitive profiling of miRNAs.

The technique employs an array of diamond nanoneedles that are functionalized with size-dependent RNA-binding-proteins, working as the "fishing rods" to directly pull multiple miRNA targets out of cytoplasm while keeping the cells alive, thus enabling quasi-single-cell miRNA analysis. Each nanoneedle works as a separated reaction chamber for parallel in-situ amplification, visualization and quantification of miRNAs as low as femtomolar, which is sufficient to detect miRNAs of a single copy intracellular abundance. The technique is also specific to single-nucleotide variation by discriminating closely-related miRNA sequences. Using the new RNA profiling technique, the research team analyzed the temporal miRNA transcriptome over the differentiation of embryonic stem cells (ESCs) towards motor neurons. The combinatorial miRNA expression patterns derived by inCell-Biopsy identifies defined cell subpopulations resulted from ESC differentiation, and also reveals the dynamic evolution of cellular heterogeneity.

The findings were recently published in the scientific journal Science Advances, titled "High-throughput intracellular biopsy of microRNAs for dissecting the temporal dynamics of cellular heterogeneity".

Press coverage:
(10 Jun 2020) Intracellular Biopsy Technique For Fast MicroRNAs Profiling In Living Cells [Scienmag]
(10 Jun 2020) Intracellular Biopsy Technique For Fast MicroRNAs Profiling In Living Cells [Phy.org]