Fig. 2 |. Development of doxycycline responsive adaptamers.
(a) Schematic of doxycycline adaptamer with variable transducer sequence, stem length, and root length. (b) Results of pooled in vitro RNA editing screen in HEK293T cells with 80μM doxycycline identifying several top candidates. ADAR reporter RNA editing in blue didn’t change in response to doxycycline. (c) Individual validation of doxycycline responsive adaptamers verified their functionality. (d) Doxycycline adaptamers were responsive to changes in ADAR1 protein levels. The same shRNA cell lines were used as in tetracycline adaptamer shRNA experiment (e) Cotransfection of adaptamers demonstrated orthogonality with the tetracycline controlled adaptamer only responsive to tetracycline and the doxycycline adaptamer only responsive to doxycycline. Error bars represent mean ± S.E.M (n = 3), and individual dots represent biological replicates. Percent expression is calculated as mean fluorescence intensity or luminescence normalized to the no-stop-codon control with tetracycline.