MOESM3 of EFTUD2 gene deficiency disrupts osteoblast maturation and inhibits chondrocyte differentiation via activation of the p53 signaling pathway

Additional file 3: Figure S2. Targeting site and efficacy of eftud2-targeted TALEN mRNAs. We also prepared three TALENs to construct the knockout zebrafish model, and the second one (T2) was the most effective for in vitro screening. A, B: The TALEN (T2) was designed at the first exon, which is presented in capital letters. C: In vitro efficacy was evaluated based on relative luciferase activity in TALEN-transfected Hek293T cells and the negative control. D: PCR products of eftud2 containing the target sequence could be digested by the T7E1 enzyme, in which the product from the mutant zebrafish was cleaved into two fragments, whereas that from WT zebrafish was intact. E: Sequencing results of F0 generation showed mixed signals from the target site, which may predict the combination with TALEN mRNA and the eftud2 gene.