Generation of gene disruptions by transcription activator-like effector nucleases (TALENs) in Xenopus tropicalis embryos
1 School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, P. R. China
2 Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, P. R. China
3 Department of Biology, South University of Science and Technology of China, Shenzhen 518055, P. R. China
4 Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen 518057, P. R. China
Cell & Bioscience 2013, 3:21 doi:10.1186/2045-3701-3-21Published: 10 May 2013
Transcription activator-like effector nucleases (TALENs) are novel engineered DNA nucleases, and have been proven to be effective for gene specific targeting in various species. Recently we reported gene disruptions in Xenopus embryos by using TALENs. Here we summarize the protocol that is used in our studies for gene disruption. This protocol covers selection of TALEN targeting sites, TALEN assembly with a modified Golden Gate method, and injection of TALEN mRNAs into Xenopus tropicalis embryos. We also provide details for detection of somatic and germ line transmitted mutations. And finally, we briefly describe establishment of knockout Xenopus lines. This protocol will facilitate broader applications of TALENs in studies of Xenopus biology.