Extended Data Fig. 3: Dual signal recording using the GFP** reporter.
From: Scalable biological signal recording in mammalian cells using Cas12a base editors
a, Schematic showing the GFP** dual recording reporter, which expresses GFP upon the activity of two distinct GFP* guides removing two stop codons. b, Left, representative histograms of HEK cells 3 days post-transfection with GFP**, base editor, and combinations of U6-crGFP*. Right, quantification of mean GFP for 3 independent replicates. c, Modified lentiviral constructs used to transduce two inducible guides into cells and select with a single antibiotic, through use of split hygromycin. d, Representative histograms of GFP** HEK cells stably transduced with NFKB and TRE3G inducible guides shown in c and transfected with base editor, 3 days posttransfection and stimulation with TNFα/dox. e, Quantification of mean fluorescence (left) and percentage GFP + (right) of GFP** HEK cells stably transduced with NFKB and TRE3G inducible guides and transfected with base editor, 3 days post transfection and stimulation with TNFα/dox. All data shown for 3 independent replicates. f, Quantification of A→G mutations at each targeted stop codon in the GFP** locus as measured by MiSeq. Data are shown for 3 independent replicates.
