TALEN constructs were assembled by the Hope Center for Neurological Disorders at Washington University using the Golden Gate TALEN assembly protocol 53 (link). The repeat variable diresidue sequences for the left and right TALENs were as follows: Left: NN NG NG NG HD NG HD HD HD NG NI NN NI HD NI NI NG and Right: NI NI HD NG NN NG NI NG HD NN HD NI NN HD HD NI NG HD NI NG. Synthetic TALEN mRNA was then transcribed using the mMESSAGE mMACHINE® SP6 ULTRA kit (Ambion), and RNA from each arm was combined and injected into 1-cell stage WT embryos (AB) at 100 pg quantities. To recover germ-line transmitted mutations, injected founders (F0s) were raised to adulthood, outcrossed to WT AB partners, and genomic DNA was extracted from individual F1 embryos for PCR amplification and restriction digest analysis of the targeted region. Genomic DNA from F1 embryos that showed disruption of the target site was then cloned using the TOPO® TA Cloning® kit (Invitrogen) and sequenced using Sanger sequencing.