Fig. 2: Identification of a lead candidate, E8, from a tumoricidal oligo library.

Cell types are denoted in the top left corner of plots. a Sequence abundance plot from a representative response-directed in vitro evolution run. The plot shows a random sample of 1,000 sequences out of the 10,000 most abundant sequences in each round (traces are shown from their first appearance in the data) with the 10 most abundant ones highlighted in color. These were synthesized and screened to find candidates. b A representative screening to highlight effective oligos. The response was measured as the ability to induce significant cas-3/7 activation in the population compared with vehicle. V, vehicle; R, random oligonucleotide; 1–10, oligo IDs (E1, E2, …, E10). c Simulated structure of E8 (see Supplementary Note 5 for derivation). d Selectivity of E8 to TNBC9 cells (blue) over the negative target cells, MCF10A (red). The response was measured by a cell viability count assay. STA, staurosporine; PAC, paclitaxel; Random, random oligonucleotide. e The effect of E8 (100 μM) on MDA-MB-231 cells. Rnd, random oligonucleotide; PAC, paclitaxel. f The dose-response curve for E8 (blue) and PEGylated (PEG)-E8 (green), showing persistence of effect in the modified oligo. The two curves are statistically indifferent. g The effect of E8 (100 μM) on TNBC cells in mouse serum. Ve, vehicle; STA, staurosporine. Chemotherapeutic controls were given at equivalent concentrations to E8.