Figure 5
From: A living biobank of canine mammary tumor organoids as a comparative model for human breast cancer
CMT organoids can be genetically modified with the CRISPR/Cas9 system. (a) GFP was introduced into ORG by lentiviral transduction at the indicated multiplicities of infection (MOI). GFP expression was analyzed by flow cytometry 3 days after transduction. For MOI 1, Mean ± SD = 12.6 ± 4.2%, for MOI 4, Mean ± SD = 35.3 ± 6.6%. Two independent experiments that were conducted with three different organoid lines are presented, demonstrating robust transduction efficiency. (b) Frequency of frameshift indels in organoids ORG-25-C1 modified by CRISPR/Cas9 using a gRNA targeting vimentin (gVIM-1 and gVIM-2), compared to the non-targeting (NT) gRNA. TIDE analysis two passages after transduction. (c) Representative images of H&E stainings and immunohistochemistry of VIM in organoids ORG-25-C1 modified by CRISPR/Cas9 using a gRNA targeting vimentin (gVIM-1 and gVIM-2), compared to the non-targeting (NT) gRNA. Embedding four passages after transduction. Scale bar, 50 μm.
