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. 2017 Feb 24;8(17):27839–27853. doi: 10.18632/oncotarget.15711

Figure 1. TALENs-induced DLEU1 knockdown and DLEU1 stably overexpressing Raji cell line.

Figure 1

(A) A diagram showing the targeting of the DLEU1 locus by three different TALENs and TALE repeat arrays are shown with the repeat-variable di-residue (RVD) underlined and represented by small colored rectangles boxes. Two monomeric TALENs (L and R) are required to bind the DLEU1 target site to cleave DNA by fused FokI. TSS, transcription start site. (B) Single clones with TALEN1 and 3-induced DLEU1 locus disruption (22,843bp deletion, boxed with red dot line) were isolated. (C) Comparison of expression of DLEU1 mRNA between WT control (WT) and DLEU1 knockdown single clones T13-2-2, T13-2-4, T13-2-14 and T13-2-16 by qRT-PCR. Data are presented as mean ± SD (n = 3, paired t test) and p-value are displayed on plot. (D) Diagram of DLEU1 gene and ORF (78 a.a). Empty GFP vector (GFP) and GFP fused DLEU1 (GFP-DLEU1) constructs are shown. ATG, start codon; TAG, stop codon. (E) GFP and GFP-DLEU1 fusion plasmids were stably transfected into Raji cells, and then western blotting (F) and qRT-PCR (G) were performed. Data are presented mean ± SD of triplicates (paired t test). *p < 0.005.