Extended Data Fig. 4: Multiplexed recording of endogenous transcripts from sugar catabolic pathways.
From: RNA recording in single bacterial cells using reprogrammed tracrRNAs
(a) Recording the rhaB, xylA and xylF transcripts in E. coli. The expression of each gene is induced by the indicated sugar. The left indicates the coloring scheme used in b and f. (b) Bulk sequencing of recorded rhaB expression induced by L-rhamnose. (c) Detecting L-rhamnose-induced population heterogeneity by flow cytometry analysis with a fluorescent transcriptional reporter. Each dot represents the mean of independent experiments starting from three separate colonies. The area of each dot scales with the fraction of cells in that population. (d) and (e) Recording of rhaB expression transiently induced with L-rhamnose. (f) Bulk sequencing of recorded xylA and xylF expression induced by D-xylose. See a for the bar coloring. Expression of the crRNA controls was driven by a constitutive promoter J23119, where editing is expected even in the absence of D-xylose. (g) Constructs for multiplexed recording of xylA and xylF expression. The DNA targets are encoded in different configurations and are spaced by different distances. (h) Multiplexed recording of xylA and xylF expression in the presence of D-xylose. Solid dot, presence of the Rptr; hollow dot, absence of the Rptr. Values in b, d, e, f and h represent the mean and standard deviation of independent experiments starting from three separate colonies.
