Single-Cell\nMultiplexed Signal Amplification Strategy\nBased on Catalytic Hairpin Self-Assembly and CRISPR/Cas12a for Exploring\nthe Relationship between lncRNA HOTAIR and miRNA-122 in Individual\nHepatocytes

The long noncoding RNA (lncRNA) HOTAIR has been shown\nto act as\nan oncogene in a variety of cancers, including hepatocellular carcinoma\n(HCC). MicroRNA-122 (miR-122) is a key liver-specific miRNA that is\nfrequently inhibited in HCC and is associated with poor prognosis.\nHowever, a potential relationship between HOTAIR and miR-122 in individual\nhepatocytes has not been explored. To this end, we propose here an\nintracellular catalytic hairpin self-assembly-CRISPR/Cas12a tandem\nmultiplexed signal amplification strategy for the simultaneous quantification\nof HOTAIR and miRNA-122 in a single hepatocyte. We applied this method\nto analyze both normal HL-7702 liver cells and HepG2 HCC cells, and\nfound that HL-7702 cells contained large amounts of miRNA-122, while\nthe content of miRNA-122 in HepG2 cells was low. However, the level\nof HOTAIR in HepG2 cells was much higher than that in HL-7702 cells,\nconfirming the overexpression of HOTAIR in HCC cells. We achieved\nthe simultaneous absolute quantification of HOTAIR and miRNA-122 in\nsingle cells, providing an important method to study the relationships\nbetween these two RNA molecules in individual cells.