Simple and efficient custom transcription activator-like effector gene synthesis via twin primer assembly

BioTechniques

Huan Wang¹  Mengyao Qu²  Yan Yu²  Yushang Zhao³  Song Wang³  Yanbing Zhu³  Yuhualei Pan⁴ 

[1] Rehabilitation, Beijing 100068, China;Translational Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China;;1Experimental &;2Institute of Rehabilitation Science of China, China Rehabilitation Research Center, Beijing Key Laboratory of Neural Injury &

关键词: genome editing;    TALE;    TALEN;    TET1;    twin primer;   

DOI  :  10.2144/btn-2020-0130

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown