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. 2020 Aug 17;31(15-16):839–851. doi: 10.1089/hum.2020.053

Figure 5.

Figure 5.

Expression of miniFXN constructs using AAV vectors in human cells. (A) Constructs miniFXN 7 and 8 were cloned into a pAAV vector (Addgene 27970) in two formats: with and without Flag tag (-F). Importantly, pAAV constructs harbor polyA (pA) signals. (B) All four constructs were transfected to HEK293T cells to verify FXN expression. Western blots were performed with a frataxin-specific antibody; FXN [en] indicates migration of the endogenous frataxin; C—control, untransfected cells, M—molecular-weight protein marker. No statistically significant difference in FXN expression between these constructs was observed. (C) AAV9 vectors were prepared using constructs miniFXN 7 and 8 containing the Flag tag and transduced to HEK293T cells at 10e5 viral genomes (vg)/cell. Expression of frataxin was analyzed by Western blot. FXN [AAV] indicates AAV-expressed FXN. C—control, untransduced cells. M—molecular-weight protein marker.