Fig. 4
From: Programmable sequential mutagenesis by inducible Cpf1 crRNA array inversion
Pooled sequential mutagenesis to model acquired resistance to immunotherapy. a Schematic of the experimental approach for pooled sequential mutagenesis using Cpf1-Flip. Following restriction digest, a library of FlipArrays is cloned into the base vector. In each FlipArray, the first crRNA targets a tumor suppressor (Nf1), while the second crRNA targets a panel of putative immunomodulatory factors. Cre-mediated inversion induces expression of the second crRNA. b Dot plot detailing the total variant frequencies at the crNf1 target site in uninfected cells, 14 days after FlipArray transduction (−Cre), and 28 days after FlipArray transduction (+Cre). c Dot plot detailing the total variant frequencies at the second crRNA target sites (Fasl, Ido1, Jak2, Lgals9, B2m, and Cd274) in uninfected cells, 14 days after FlipArray transduction (−Cre), and 28 days after FlipArray transduction (+Cre). All error bars are mean ± s.e.m (n = 3 cell replicates for all conditions).
