(A) Two individual Nf1 sgRNAs accelerated subcutaneous tumor growth. The tumors (n = 4) were derived from p53; Myc; Cas9 cells (Ctrl) infected with sgNf1.1 and sgNf1.4, independently. Error bars, mean ± s.e.m. The inset shows that sgNfl unregulated the phosphor-Erk (pErk) with Hsp90 as a loading control. ***, P < .001. Error bars, mean ± s.e.m.
(B) Schematic of hydrodynamic delivery of plasmids encoding Myc transposon (Tn) and Cas9/sgRNA (targeting Nf1, or GFP control) into liver-specific p53-knockout mouse. Representative images of livers from mice treated with sgGFP control (left) or sgNf1 (right) at 1 month after injection are shown. Arrows denote tumors.
(C) Quantification of tumor number per mouse (n = 4 mice) shown in (B).
(D) Sequences of Nf1 sgRNA target sites from representative liver tumors showing indel mutations and the fraction of reads for each.