Abstract
We report gene targeting via homologous recombination in zebrafish. We co-injected fertilized eggs with transcription activator–like effector nuclease mRNAs and a donor vector with long homologous arms targeting the tyrosine hydroxylase (th) locus, and we observed effective gene modification that was transmitted through the germ line. We also successfully targeted two additional genes. Homologous recombination in zebrafish with a dsDNA donor expands the utility of this model organism.
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References
Capecchi, M.R. Nat. Rev. Genet. 6, 507–512 (2005).
Doyon, Y. et al. Nat. Biotechnol. 26, 702–708 (2008).
Meng, X., Noyes, M.B., Zhu, L.J., Lawson, N.D. & Wolfe, S.A. Nat. Biotechnol. 26, 695–701 (2008).
Huang, P. et al. Nat. Biotechnol. 29, 699–700 (2011).
Sander, J.D. et al. Nat. Biotechnol. 29, 697–698 (2011).
Huang, P., Zhu, Z., Lin, S. & Zhang, B. J. Genet. Genomics 39, 421–433 (2012).
Wang, D. et al. Proc. Natl. Acad. Sci. USA 104, 12428–12433 (2007).
Huang, P. et al. Nucleic Acids Res. 41, e36 (2013).
Chen, F. et al. Nat. Methods 8, 753–755 (2011).
Bedell, V.M. et al. Nature 491, 114–118 (2012).
Cui, X. et al. Nat. Biotechnol. 29, 64–67 (2011).
Shukla, V.K. et al. Nature 459, 437–441 (2009).
Bibikova, M., Beumer, K., Trautman, J.K. & Carroll, D. Science 300, 764 (2003).
Urnov, F.D. et al. Nature 435, 646–651 (2005).
Doyle, E.L. et al. Nucleic Acids Res. 40, W117–W122 (2012).
Cermak, T. et al. Nucleic Acids Res. 39, e82 (2011).
Elliott, B., Richardson, C., Winderbaum, J., Nickoloff, J.A. & Jasin, M. Mol. Cell Biol. 18, 93–101 (1998).
Alexander, J., Stainier, D.Y. & Yelon, D. Dev. Genet. 22, 288–299 (1998).
Paul, P.K. et al. J. Cell Biol. 191, 1395–1411 (2010).
Acknowledgements
We thank M. Veldman, Z. Tehrani and J. Ear for carefully proofreading and editing the manuscript; S. He, X. Yang, X. Wang and J. Chen for help in founder screening; Y. Shen, Y. Gao and J. Zhang for technical support and lab management; Y. Jia, J. Chen, X. Yang and H. Cui for zebrafish maintenance; Y. Li, X. Yang and K. Sampath for help in establishing the Southern blot method; and J. Liu for help in the methylation analysis. This work was supported by grants from the 973 program (2012CB945101, 2011CBA01000), National Natural Science Foundation of China (31110103904, 30871418, 30730056) and US National Institutes of Health to S.L. (DK54508, DK084349).
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Y.Z., X.T., B.Z. and S.L. conceived of and designed experiments; Y.Z. assembled TALENs. Y.Z. and Z. Li performed microinjections; Y.Z., Z. Li and R.P. built donor constructs; Y.Z., X.T., D.L., R.P., Z.W. and Q.W. carried out the PCR diagnosis; D.L., Z.W. and P.H. performed Southern blot analysis; Y.H. and Z. Luo performed the experiments at the smad5 locus. Y.Z., X.T., Z.Z., B.Z. and S.L. wrote the manuscript.
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Zu, Y., Tong, X., Wang, Z. et al. TALEN-mediated precise genome modification by homologous recombination in zebrafish. Nat Methods 10, 329–331 (2013). https://doi.org/10.1038/nmeth.2374
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DOI: https://doi.org/10.1038/nmeth.2374
