GSM8290551: KA951; Homo sapiens; RNA-Seq
In vivo assays were performed similarly to in vitro overexpression assays. However, only constructs for OVOL2, HNF1A, mCherry and the OVOL2-HNF1A combination were used in vivo. These viral constructs were transduced into KP4 cells in arrayed fashion (approx. MOI 1-2), followed by puromycin selections and pooling the samples together in the following ratios based on their in vitro growth rate (OVOL2 50%, HNF1A 16.6%, mCherry 16.6% and OVOL2-HNF1A 16.6%). Pooled cells were then injected into athymic mice models (3 mice in total). 14 days later the animals were sacrificed followed by scRNA-seq for the pooled samples. Similarly, as with the pooled in vitro assays, the ORFs for any given cell were identified by using ORF barcodes and specific primers for amplifying ORF barcodes (same as with in vitro assays) from the amplified cDNA. M2RTTA was not used for in vivo overexpression assays.