Extended Data Fig. 9: In vivo CRISPR-KO of Fosl1, Junb, and Klf5 block tumorigenesis driven by Krasmut and TSG KO.
a, Initial infection rates of pancreatic cells determined by flow cytometry five days after injection of the indicated sgRNA-expressing AAV-Cre-sgRNA viruses in LSL-Cas9-eGFP mice. b, H&E sections of pancreases from mice injected with AAV-Cre-sgRNA on D18, images representative of two sgRNAs are shown. Results for Klf5 and Rosa are representative of two independent experiments, while Fosl1 and Junb KO was performed once, with two independent sgRNAs. Scale bars, 100um. c, Relative efficiency of LSL cassette removal at the LSL-KrasG12D allele in eGFP-positive cells on D18 after AAV-Cre-sgRNA injection, evaluated by PCR from genomic DNA obtained from FACS-sorted cells. d, Efficiency of CRISPR/Cas9-induced deletion/insertions at the indicated sgRNA-targeted sites. To calculate this, genomic DNA was isolated from eGFP-positive cells on D18 after AAV-Cre-sgRNA injection, followed by PCR amplification of the targeted region. PCR products were subjected to Sanger sequencing, and the resulting trace files were analyzed using the Tracking of Indels by Decomposition (TIDE) algorithm (https://tide.nki.nl/). e, Normalized ATAC-seq intensity at K-LOCKEDOPEN sites after infection with the indicated AAV-Cre-sgRNA compared with sgROSA controls. P-values calculated by Kolmogorov–Smirnov test, two sided. ****, p<1x10-5, **, p<.001.
