Wang S(1), Yu Y(2), Pan Y(1), Wang H(1), Zhao Y(1), Qu M(1), Zhu Y(1). Author information:
(1)Experimental & Translational Research Center, Beijing Friendship Hospital,
Capital Medical University, Beijing 100050, China.
(2)Institute of Rehabilitation Science of China, China Rehabilitation Research
Center, Beijing Key Laboratory of Neural Injury & Rehabilitation, Beijing
Transcription activator-like effector (TALE) nucleases (TALENs) efficiently recognize and cleave DNA in a sequence-dependent manner. However, current TALE custom synthesis methods are either complicated or expensive. Here we report a simple and low-cost method for TALE construct assembly. This method utilizes the denaturation/reannealing nature of double-stranded DNA to create a unique single-stranded DNA overhang for proper ordering of TALE monomers in an engineered multimer. We successfully synthesized two TALEN pairs targeting the endogenous TET1 locus in human embryonic kidney cells and demonstrated their editing efficiency. Our method provides an alternative simple, low-cost method for effective TALEN assembly, which may improve the application of TALE-based technology.
Having over 250 Research scholars worldwide and more than 400 articles online with open access.