Mutagenesis in Xenopus and Zebrafish using TALENs.

Yun Liu, Hui Zhao, Christopher H K Cheng

Methods in molecular biology (Clifton, N.J.) 2016

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Transcription activator-like effector nucleases (TALENs) have been proven to be effective for gene specific targeting across species. Here we describe the validated protocol for TALEN assembly as well as methods for generating gene knockout animals of Xenopus and zebrafish. This protocol covers selection of TALEN targeting sites, TALEN assembly with a modified Golden Gate method, injection of TALEN mRNAs into Xenopus and zebrafish embryos as well as the detection of somatic and germ-line transmitted mutations. Finally, the establishment of knockout Xenopus and zebrafish lines is also described. This protocol will facilitate broader applications of TALENs in developmental biology.

Citation

Yun Liu, Hui Zhao, Christopher H K Cheng. Mutagenesis in Xenopus and Zebrafish using TALENs. Methods in molecular biology (Clifton, N.J.). 2016;1338:207-27