Fig. 2.

Detection of ecDHFR DD-fused RESCUE system expression induced by TMP. a Schematic representation of the engineered ecDHFR DD-fused RESCUE expression plasmids. ecDHFR DD was fused to the C-terminal of mCherry. b Illustration of the mechanism by which conditionally destabilized ecDHFR DD regulates dRanCas13b, ADAR2, and mCherry expression. ADAR2 and mCherry are fused with the ecDHFR DD. The TMP prevents the destabilization and degradation of dRanCas13b, ADAR2, and mCherry. c dRanCas13b, ADAR2, and mCherry expression in RESCUE- and ecRESCUE-transfected 293T cells with or without TMP. Without TMP (24 h): at 48 h post-transfection without TMP; + TMP: at 24 h post-TMP treatment; +  − TMP (24 h): at 24 h post-TMP removal; +  − TMP (48 h): at 48 h post-TMP removal. d Red optical density statistics of mCherry in RESCUE- and ecRESCUE-transfected 293T cells. RESCUE and ecRESCUE: at 48 h post-transfection without TMP; + TMP: at 24 h post-TMP treatment; − TMP (24 h): at 24 h post-TMP removal; − TMP (48 h): at 48 h post-TMP removal. Values are shown as mean ± SEM (n = 3). Different superscript letters indicate significant differences as follows: *P < 0.05; **P < 0.01; ***P < 0.001; ns indicate there is no significance