The first exon sequences of Dll gene were scanned for potential TALEN target sites, which were identified using the TALEN Targeter program at https://tale-nt.cac.cornell.edu/node/add/talen . The following parameters were used: 1) spacer length: 12-16; 2) repeat array length of 15-20; 3) G substitute: NN; 4) Filter Options: Show all TALEN pairs (include redundant TALENs); 5) Streubel et al. guidelines: On. The Platinum Gate TALEN construction system described in Sakuma et al. [14 (link)] was used to assemble two pairs of TALENs, Dll_ A and Dll_ B (Figure 1 ), using the Platinum Gate TALEN Kit (Addgene, cat#1000000043). Assembled RVD repeats were as follows: HD NN NI HD NN NI HD NN HD HD HD NI NI NG NG HD for the left Dll_A TALEN, NN NG NG NN NG NG NN HD NG NN NN NG HD NN HD NI NI for the right Dll_A TALEN, HD NI NG HD HD NN NN NI HD NI NN HD NI NN HD NI NN for the left Dll_B TALEN, and NN HD NN NN HD NN NN HD HD NN HD NG NN HD NI NN HD for the right Dll_B TALEN. ptCMV-153/47-VR vectors (Addgene, included in the Platinum Gate TALEN Kit) were used as destination vectors.
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