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Mammalian Expression> | … To prepare cDNA encoding Csy4, human-codon-optimized Csy4 was synthesized by GenScript (Piscataway, NJ, USA). The cDNAs encoding Csy4RS-ZsG and Csy4 were inserted into the NheI/XhoI and EcoRI/XbaI sites of pLV.PGb.FtR, respectively, to yield pLV.PC.FCRS-Z. … | Get A Quote |
MicroRNAs (miRNAs) are small noncoding RNAs that regulate complex gene expression networks in eukaryotic cells. Because of their unique expression patterns, miRNAs are potential molecular markers for specific cell states. Although a system capable of imaging miRNA in living cells is needed to visually detect miRNA expression, very few fluorescence signal-on sensors that respond to expression of target miRNA (miR-ON sensors) are available. Here we report an miR-ON sensor containing a bidirectional promoter-driven Csy4 endoribonuclease and green fluorescent protein, ZsGreen1, for live-cell imaging of miRNAs with post-transcriptional feedback control. Csy4-assisted miR-ON (Csy4-miR-ON) sensors generate negligible ... More