Extended Data Fig. 7: CaTCH isolation of specific clones allows their phenotypic characterization.
From: Isolating live cell clones from barcoded populations using CRISPRa-inducible reporters
a, NGS of CaTCH isolated clones identified in the in vivo experiment (see Fig. 2d). Shown is the BC composition after CaTCH isolation of the indicated BC from the starting population (treatment-naïve and depleted, Fig. 2e) or from tumor-derived RAFi/MEKi-resistant cell lines. BCs > 1% of the sample are highlighted individually in color, BCs < 1% were totaled in white bars. The correct, targeted BC within each sample is shown at the bottom section of every bar, in its respective color code from Fig. 2d, e. b, BC composition of different isolations of BC14559 or BC43158. Both BCs were always identified together, indicated in the respective BC color. The BC which was targeted for isolation is indicated below each bar. c, Tumor growth curves and survival curves of in vivo RAFi/MEKi treatment response of three randomly single cell sorted CaTCH-barcoded clones (C1, C2, C3). Sorted clones were transduced with a barcode-unspecific CaTCH reporter activating sgRNA and sorted for activated GFP signal before injection. Growth curves displayed as mean ± s.e.m. Statistical analysis on survival curves was performed by two-sided Log-rank (Mantel-Cox) test, comparing individual clones on treatment to untreated. *P < 0.05, ns=non-significant. n = tumors, two tumors per mouse, C1: UT n = 6, RAFi/MEKi n = 8; C2: UT n = 6, RAFi/MEKi n = 8; C3: UT n = 4, RAFi/MEKi n = 8). d, Survival curves corresponding to the tumor growth curves from Fig. 2f. Statistical analysis was performed by two-sided Log-rank (Mantel-Cox) test, comparing individual clones on treatment to untreated. *P < 0.05, **P < 0.01, ns=non-significant. n=mice. BULK: TNUT n = 3, TNRAFi/MEKi n = 5; BC121: TNUT n = 3, RUT n = 3, TNRAFi/MEKi n = 5, RRAFi/MEKi n = 5; BC952: TNUT n = 3, RUT n = 3, TNRAFi/MEKi n = 4, RRAFi/MEKi n = 5; BC2487: TNUT n = 3, RUT n = 3, TNRAFi/MEKi n = 4, RRAFi/MEKi n = 5; BC13: TNUT n = 3, TNRAFi/MEKi n = 4; BC2721: TNUT n = 3, TNRAFi/MEKi n = 5; BC2646: TNUT n = 3, RUT n = 2, TNRAFi/MEKi n = 5, RRAFi/MEKi n = 3; BC14559: TNUT n = 3, RUT n = 3, TNRAFi/MEKi n = 3, RRAFi/MEKi n = 5.
