![]() ![]() Furthermore, we generated genetically engineered mice harbouring circRNA expression constructs. We further demonstrated ectopic circRNA expression in a hepatocellular carcinoma mouse model upon circRNA transposon delivery via hydrodynamic tail vein injection. We demonstrated that circRNA expression constructs can be delivered to cultured cells via transposons, whereas lentiviral vectors have limited utility for the delivery of circRNA constructs due to viral RNA splicing in virus-producing cells. Here we report the development and characterization of genetic tools enabling stable circRNA overexpression in vitro and in vivo. While mammalian expression plasmids enable transient circRNA overexpression in cultured cells, most cell biological studies require long-term ectopic expression. Overexpression studies in particular suffer from the lack of efficient genetic tools. Standard gain-of-function and loss-of-function approaches to study gene functions have significant limitations when studying circRNAs. circRNAs are broadly expressed and contribute to biological processes through a variety of functions. Circular RNAs (circRNAs) are a class of non-coding RNAs featuring a covalently closed ring structure formed through backsplicing.
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