Genome Modification of Pluripotent Cells by Using Transcription Activator-Like Effector Nucleases (TALENs).

Amir Taheri-Ghahfarokhi, Luis F Malaver Ortega, Huseyin Sumer

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)

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 modifi ed. Here, we describe a modifi ed 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.
Original languageEnglish
Title of host publicationCell Reprogramming
Subtitle of host publicationMethods and Protocols
EditorsPaul J Verma, Huseyin Sumer
Place of PublicationNew York NYJ USA
PublisherHumana Press
Pages253-268
Number of pages16
Edition1st
ISBN (Print)9781493928477
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology: Springer Protocols
PublisherSpringer
Number1330
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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