Figure 4.

Increased Presence of Editing Sites on Chromosome One and Increased Density of Editing around the MBS Reporter Locus

(A) Schematic of the reporter locus on chromosome 1p36. Lac operator repeats (LacO) are present upstream of 96 repeats of the tet response element (TRE); when bound by reverse tet transactivator, the cytomegalovirus (CMV) promoter is inactive. Upon doxycycline induction the RNA containing the MBS array becomes actively transcribed (right arrow) at which point, MCP-ADAR (blue circles) can bind to the RNA at the site of transcription.

(B) Reporter MBS gene, focusing on the MBS array showing (from top to bottom) the following. MS2 sites at genomic locus (ground truth): blue horizontal bars represent the known location of the MS2 stem loops. MCP-TRIBE sites: editing events as indicated by dark blue bars where height corresponds to the average editing percentage across both replicates at that nucleotide (scale to right). Light blue shading indicates location of the stem-loop nucleotides.

(C) Bar graph showing number of editing sites (y axis) along chromosome one (x axis, 10-Mb bins) at 24 h post-doxycycline induction.

(D) Bar graph showing number of editing sites (y axis) along chromosome one (x axis, 10-Mb bins) at 48 h post-doxycycline induction.

(E) Bar graph showing number of genes edited (y axis) organized by chromosome of origin (x axis). Relative number of editing sites calculated by normalizing genes edited per chromosome to the total number of genes edited in the experiment. Chromosomes are colored by experiment, and colors correspond to (B). Gray bars representing cells with no doxycycline induction (low expression), blue bars represent 24 h of doxycycline induction, and red bars represent 48 h of doxycycline induction.