Over Expression of Specific Transcription Factors to Boost FVIII Expression

Background Main source of FVIII production and secretion is the liver sinusoidal endothelial cells, whereas other endothelial cells also express and secrete FVIII but to a less extent. This variability in expression in different endothelial cells could be caused by epigenetic regulation and/or by differential expression of transcription factors binding to the promoter region. To address the second point, we previously used TRANSFAC database to identify in silico potential transcription factors binding to the FVIII promoter and determined their expression levels in different endothelial cells. We identified a group of TFs that are relatively overexpressed in high expressing cells.

Aim In this pilot study, we wanted to evaluate the effect of over expression of our identified TFs on FVIII expression levels. For this purpose, we choose to start with three TFs: HNF1A, EBF3, and USF1.

Materials and Methods We transfected pcDNA3 vectors containing the TFs of our choice into HEK293 cells using two transfection methods: (A) the lipofection and (B) the nucleofection; the F8 expression level was monitored by real-time PCR.

Results FVIII expression levels were higher for HNF1A than for EBF3 and USF1; however, all appears close to the mock control (which was also variable) but where higher than the untransfected control. This effect could be specific for HEK cells used in this experiment.

Conclusion The result of this pilot study shows that introducing specific TFs can modulate the F8 expression.

Fig. 2

^*These authors equally contributed to this study.

Publication History

Article published online:
13 November 2020