Extended Data Fig. 8: Utilizing two parallel guides for more detailed record of input signal characteristics.
From: Scalable biological signal recording in mammalian cells using Cas12a base editors
a, Schematic showing two input signals with distinctive patterns but the same area under the curve. b, Promoter engineering strategy used to reduce the sensitivity of NFKB-responsive promoter, to generate two inducible guide constructs that respond differently to input TNFα signal. c, Mean BFP fluorescence of HEK cells 3 days post-transfection with NFKB promoter variants and stimulation with TNFα. Left, raw mean BFP values are plotted. Right, mean BFP values normalized to the maximum of each respective promoter are plotted. Error bars indicate standard deviation of 3 independent replicates. d, Quantification of %A→G conversion as measured by MiSeq for HEK cells transfected with base editor, 5X-NFKB-crAAVS1_4, and 2X-NFKB-crAAVS1_8. One day after transfection, cells were stimulated with either 100 ng/mL TNFα for 0.25 hours, or 0.5 ng/mL TNFα for 50 hours. All data shown for 3 independent replicates. Statistical analysis was performed using a two-sided t test without adjustment for multiple comparisons.
