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. 2019 Sep 16;2(5):e201900358. doi: 10.26508/lsa.201900358

Figure S1. Generation of a TDP-43 KO cell line.

Figure S1.

(A) Alignment of WT sequence and mutant sequence containing a 7-bp deletion at the Cas9 cut site in TDP-43 exon 3 (region coding for RRM1). Regions recognized by gRNA#1 and gRNA#2 are highlighted in yellow and green, respectively. Only gRNA#1 was used for the generation of the TDP-43 KO clonal line. (B) Alignment of WT sequence and mutant sequence containing a 1-bp insertion at the Cas9 cut site in TDP-43. (C) Summary of the mutations detected in the TDP-43 KO cell line by sequencing of the region of genomic DNA targeted by the TDP-43 sgRNA. (D) Immunoblotting results from WT and TDP-43 KO cells reveal the absence of both full-length TDP-43 protein as well as of any fragments arising from the mutant alleles. (E) Immunoblotting results from WT and TDP-43 KO cells using an antibody raised against the N terminus of TDP-43 showing absence of TDP-43 fragments in KO cells.