biostudies-literatureGuo Ythe Outstanding Young teacher sponsored by Qing Lan Project in Jiangsu Provincethe National Natural Science Foundation of China1172-1178https://www.ebi.ac.uk/biostudies/studies/S-EPMC9648401biostudies-literatureUnknown19(1)No current RNA-targeted interference tools have been reported to simultaneously up and down-regulate different gene expressions. Here we characterized an RNA-targeted genetic regulatory strategy composed of a flap endonuclease 1 (FEN1) and specific mis-hairpin DNA probes (mis-hpDNA), to realize the orthogonal genetic regulation. By targeting mRNA, the strategy hindered the translation and silenced genes in human cells with efficiencies of ~60%. By targeting miRNA, the strategy prevented the combination of miRNA to its specific mRNA and increased this mRNA expression by about 3-folds. In combination, we simultaneously performed <i>CXCR4</i> gene knock-down (~50%) and <i>EGFR</i> gene activation (1.5-folds) in human cells. Although the functional property can be further improved, this RNA-taRNA biologyA novel strategy for orthogonal genetic regulation on different RNA targeted loci simultaneously.PMC96484018215020932071440Liu YTian KWang PQi ZGuo YZhang XXu SQiang HZhou GWang LfalseA novel strategy for orthogonal genetic regulation on different RNA targeted loci simultaneously.No current RNA-targeted interference tools have been reported to simultaneously up and down-regulate different gene expressions. Here we characterized an RNA-targeted genetic regulatory strategy composed of a flap endonuclease 1 (FEN1) and specific mis-hairpin DNA probes (mis-hpDNA), to realize the orthogonal genetic regulation. By targeting mRNA, the strategy hindered the translation and silenced genes in human cells with efficiencies of ~60%. By targeting miRNA, the strategy prevented the combination of miRNA to its specific mRNA and increased this mRNA expression by about 3-folds. In combination, we simultaneously performed <i>CXCR4</i> gene knock-down (~50%) and <i>EGFR</i> gene activation (1.5-folds) in human cells. Although the functional property can be further improved, this RNA-ta2022-01-01T00:00:00Z2022 Jan2025-04-05T12:32:08.856Z2025-04-05T12:32:08.856ZS-EPMC96484013635079010.1080/15476286.2022.2141507