Interest is increasing in transcription activator-like effector nucleases (TALENs) as a tool to introduce targeted double-strand breaks into the large genomes of human and animal cell lines. The produced DNA lesions stimulate DNA repair pathways, error-prone but dominant non-homologous end joining (NHEJ) and accurate but less occurring homology-directed repair (HDR), and as a result targeted genes can be modified. Here, we describe a modified Golden-Gate cloning method for generating TALENs and also details for targeting genes in mouse embryonic stem cells. The protocol described here can be used for modifying the genome of a broad range of pluripotent cell lines.
CITATION STYLE
Taheri-Ghahfarokhi, A., Malaver-Ortega, L. F., & Sumer, H. (2015). Genome modification of pluripotent cells by using transcription activator-like effector nucleases (TALENs). In Methods in Molecular Biology (Vol. 1330, pp. 253–267). Humana Press Inc. https://doi.org/10.1007/978-1-4939-2848-4_19
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