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: We have previously developed a cell-based aggregation model by cloning in-tandem 12 copies of the Q/N region from TDP-43 in the C-terminal of EGFP, resulting in the expression of a so-called EGFP-12xQ/N protein (16). In order to generally improve the aggregation process and to further study whether other regions in TDP-43 could contribute to it we considered to include the 12xQ/N repetitions into the C-terminus of the Flag-TDP-43 protein itself (Fig. 1A).Figure 1.
Affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), 34012 Trieste, Italy.