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Updated: Mar 29, 2026

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
Published on: February 2, 2016
Genome Modification of Pluripotent Cells by Using Transcription Activator-Like Effector Nucleases (TALENs)
Amir Taheri-Ghahfarokhi1, Luis F Malaver-Ortega2,3, Huseyin Sumer4
1Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran. taheri1362@gmail.com.
Abstract:
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.
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