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. 2023 Apr 18;6(7):e202302005. doi: 10.26508/lsa.202302005

Figure 2. Growth competition assay in mouse embryonic stem cells.

Figure 2.

(A) Schematic representation of the genetrap system when inserted into an intron. When in sense orientation, the genetrap exposes a splicing acceptor site and will be inserted into the mature mRNA, leading to a knockout of the gene of interest. When in antisense orientation, the splicing acceptor site is inactive and the genetrap will be spliced out, leading to a WT expression of the gene of interest. Flipping of the genetrap orientation is possible by expressing the Cre recombinase. (B, C, D) Growth competition assay between sister clones bearing a genetrap into the gene of interest ((B): LRP1, (C): PTAR, and (D): FBXW11). Sister clones with WT phenotype are in grey, and sister clones with knockout phenotype are in black (see Table 1). ∼30% of knockout cells were mixed with 70% of their WT sister clone, and infected with RVFV MP-12 at an MOI of 5. The ratio between both sister clones was followed by flow cytometry. n = 2, except if the event count was below 1,000, in which case the whole data set was removed (n = 1).