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. 2023 Sep 20;34:102038. doi: 10.1016/j.omtn.2023.102038

Figure 1.

Figure 1

Rationale and scaffold screening for the building of a multiplex miRNA-based shRNA platform

(A) Effect on KD efficiency of the use of the same scaffold in tandem repeats as evaluated by flow cytometry. The KD efficiency of four different shGuide sequences (expressed as mean fluorescence intensity [MFI]) was compared in a single miR196a-2 scaffold and in the context of a 4-plex made of repeats of the miR196a-2 scaffold transduced in CAR T cells. (B) Schematic representation of the four miRNA cluster most highly expressed in T cells with an average scaffold size below 250 bases. (C) KD efficiency in CAR T cells of the miRNA scaffolds belonging to the four clusters most highly expressed in T cells with an average scaffold size below 250 bases. KD efficiency was measured by flow cytometry as MFI relative to a control without shRNA, using validated shRNA-derived sequences. Bars represent mean ± standard deviation (SD) for three independent biological replicates. Each symbol superimposed to the bars represents a different peripheral blood mononuclear cell (PBMC) donor.