Abstract: Transcription activator-like effector nuclease (TALEN) was one of the latest genome modification technologies, which was an artificially engineered hybrid protein. To ensure the specificity for the target sequence, sequences encoding transcription activator-like effectors (TALE) domains generally was so long. Therefore, construction of TALEN expression vectors was a key step in the application of TALEN. This paper introduced construction methods of TALEN expression vector, including Golden Gate, REAL, Unit Assembly, FLASH, ICA, LIC and so on, and the latest research progress. With construction methods of TALEN expression vector further enriched and popularized, this technology had broad application prospects in the biological study of gene function, crops and livestock improvement, the treatment of human genetic diseases.