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Conditional targeted genome editing using somatically expressed TALENs in C. elegans

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Abstract

We have developed a method for the generation of conditional knockouts in Caenorhabditis elegans by expressing transcription activator–like effector nucleases (TALENs) in somatic cells. Using germline transformation with plasmids encoding TALENs under the control of an inducible or tissue-specific promoter, we observed effective gene modifications and resulting phenotypes in specific developmental stages and tissues. We further used this method to bypass the embryonic requirement of cor-1, which encodes the homolog of human severe combined immunodeficiency (SCID) protein coronin, and we determined its essential role in cell migration in larval Q-cell lineages. Our results show that TALENs expressed in the somatic cells of model organisms provide a versatile tool for functional genomics.

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Figure 1: dpy-5 was conditionally edited by spatially or temporally controlled somatic TALENs in C. elegans.
Figure 2: Somatic TALENs mutated gfp in C. elegans Q-cell lineages.
Figure 3: Somatic TALENs conditionally mutated mab-5 and cor-1 and revealed the function of COR-1 in C. elegans Q-cell migration.

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Acknowledgements

We thank B. Zhang and R. Jiao for TALEN reagents and W. Zhong, D. Huangfu, L. Cai and P. Gonczy for the helpful discussion. This work was supported by National Basic Research Program of China to W.L. and G.O. (973 Program, 2012CB966800, 2012CB945002 and 2013CB945600), the National Natural Science Foundation of China to G.O., X.W., Y.C., Y.Y. and W.L. (31201009, 31201048, 31222035, 31101002, 31100972, 31171295 and 31190063), the Natural Science Foundation of Beijing to X.W. (5123045), Chinese Academy of Sciences and the Junior Thousand Talents Program of China to G.O.

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Z.C., P.Y., W.L. and G.O. designed experiments; Z.C., P.Y., X.W., Y.C., G.F., Y.Y., X.L., P.Y., Z.Z. and W.L. performed experiments; G.O. wrote the manuscript.

Corresponding authors

Correspondence to Wei Li or Guangshuo Ou.

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The authors declare no competing financial interests.

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Cheng, Z., Yi, P., Wang, X. et al. Conditional targeted genome editing using somatically expressed TALENs in C. elegans. Nat Biotechnol 31, 934–937 (2013). https://doi.org/10.1038/nbt.2674

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