TDP-43 loss of cellular function through aggregation requires additional structural determinants beyond its C-terminal Q/N prion-like domain.
Bottom Line: To our knowledge, this is the only system that achieves full functional TDP 43 depletion with effects similar to RNAi depletion or gene deletion.As a result, this model will prove useful to investigate the loss-of-function effects mediated by TDP-43 aggregation within cells without affecting the expression of the endogenous gene.These data show for the first time that cellular TDP-43 aggregation can lead to total loss of function and to defective splicing of TDP-43-dependent splicing events in endogenous genes.
Affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), 34012 Trieste, Italy.Show MeSH
Related in: MedlinePlus
Mentions: It was then of interest to use this system to investigate whether other sequences/domains of TDP-43 were contributing together with the 12xQ/N to these phenomena. To understand the contribution of the different domains of TDP-43 to induce aggregation and loss of function, we generated a variety of HEK293 stable cell lines expressing different mutants of Flag-TDP-12xQ/N (Fig. 4A). The expression of these proteins was induced by tetracycline and 72 h later TDP-43 functionality was tested by looking at the splicing of the endogenous POLDIP3/SKAR gene both at the mRNA and protein levels (Fig. 4B and C, respectively, indicated as variant 1 or variant 2).Figure 4.
Affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), 34012 Trieste, Italy.