单位：[1]Experimental & Translational Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China医技科室北京市临床医学研究所实验中心首都医科大学附属北京友谊医院[2]Institute of Rehabilitation Science of China, China Rehabilitation Research Center, Beijing Key Laboratory of Neural Injury & Rehabilitation, Beijing 100068, China
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. METHOD SUMMARY We report a simple and cost-effective transcription activator-like effector construct assembly method. This novel method is also suitable for the construction of other transcription activator-like effector fusion genes.
基金：
China Postdoctoral Science FoundationChina Postdoctoral Science Foundation [2017M620834]; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81771235]
语种：
外文
WOS：
PubmedID：
中科院(CAS)分区：
出版当年[2020]版：
大类|4 区工程技术
小类|4 区生化研究方法4 区生化与分子生物学
最新[2025]版：
大类|4 区生物学
小类|4 区生化研究方法4 区生化与分子生物学
JCR分区：
出版当年[2019]版：
Q4BIOCHEMICAL RESEARCH METHODSQ4BIOCHEMISTRY & MOLECULAR BIOLOGY
最新[2023]版：
Q3BIOCHEMICAL RESEARCH METHODSQ4BIOCHEMISTRY & MOLECULAR BIOLOGY
第一作者单位：[1]Experimental & Translational Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
共同第一作者：
通讯作者：
推荐引用方式(GB/T 7714)：
Wang Song,Yu Yan,Pan Yuhualei,et al.Simple and efficient custom transcription activator-like effector gene synthesis via twin primer assembly[J].BIOTECHNIQUES.2021,70(2):100-106.doi:10.2144/btn-2020-0130.
APA：
Wang, Song,Yu, Yan,Pan, Yuhualei,Wang, Huan,Zhao, Yushang...&Zhu, Yanbing.(2021).Simple and efficient custom transcription activator-like effector gene synthesis via twin primer assembly.BIOTECHNIQUES,70,(2)
MLA：
Wang, Song,et al."Simple and efficient custom transcription activator-like effector gene synthesis via twin primer assembly".BIOTECHNIQUES 70..2(2021):100-106
