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. 2016 Apr 19;27(6):423–424. doi: 10.1089/hum.2016.012

Figure 1.

Figure 1.

Comparison of recombination between the original pT3Ts-goldyTALEN (pT3TsgT) and the modified pKT3Ts-goldyTALEN (pKT3TsgT) vectors. (A) The original pT3TsgT vector has two lacZ sequences on opposite strands, allowing for recombination and excision of the RVD cut sites and the FokI gene. (B) The modified pKT3TsgT vector lacks the second lacZ site and has a backbone that contains the kanamycin resistance gene. (C) Transformation of pT3TsgT and plating on LB-Xgal plates with the appropriate antibiotic produces 5 times as many white colonies as blue. (D) Plating pkT3TsgT on LB-Xgal plates results in only blue colonies. (E) Comparison of targeting efficiency between TALENs assembled in the pT3TsgT and pKT3TsgT vectors. Each TALEN pair targeting either rb1 or cdh5 resulted in 100% biallelic inactivation after injection into individual embryos. +, digested; −, undigested PCR amplicons.