Extended Data Fig. 4: ADAR1 is necessary for CellREADR in 293T cells.
From: Programmable RNA sensing for cell monitoring and manipulation
a, Schematic for generating a ADAR1 knockout cell line with CRISPR/Cas9. b, Western blot analysis showing ADAR1 expression in wild-type and no expression in ADAR1 knockout cells. c, Schematics of EF1a-ChETA-tdT and READRtdT-GFP vectors (left), and ADAR1 isoform expression vectors (right). d, Both p110 and p150 ADAR1 isoforms rescued the CellREADR functionality assayed by cell conversion ratio in ADAR1 knockout cells. e–h, INF-b increased CellREADR efficiency and ADAR expression. e, Schematic of EF1a-ChETA-tdT and READRtdT-GFP vectors. f, Western blot analysis showed increased expression of ADAR1-p110 protein and induction of ADAR1-p150 isoform after interferon treatment. g, Representative FACS analysis of GFP and RFP expression with mock (left) or interferon treatment (right). h, Quantification of the READRtdT-GFP efficiency in g, which was increased by interferon treatment. Error bars in d and h are mean values ± s.e.m. n = 3, n represents the number of independent experiments performed. For gel source data, see Supplementary Fig. 2.
