Journal
METHODS
Volume 69, Issue 1, Pages 108-117Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2014.04.012
Keywords
Transcription activator-like effector nucleases; Enrichment; Mutant cells; Flow cytometry; Magnetic separation; Hygromycin B
Funding
- National Research Foundation of Korea [2011-0013568, 2011-0019357, 2013M3A9B4076544, 2008-0062287]
- Health Technology R&D Project, Ministry of Health and Welfare of Korea [HI10C1740]
- National Research Foundation of Korea [2008-0062287, 2011-0013568, 2011-0019357, 2013M3A9B4076544] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Targeted gene knockout using engineered nucleases such as transcription activator like-effector nucleases (TALENs) is a gold standard for investigating the functions of a gene of interest. Although most TALENs can cleave chromosomal DNA efficiently, the activities of designed TALENs are not always high enough to allow the efficient derivation of cells containing TALEN-driven mutations. Thus, simple methods to enrich cells containing TALEN-directed mutations would facilitate the use of TALENs. Here we describe the enrichment of such cells using surrogate episomal reporters coupled with flow cytometric sorting, magnetic separation, or hygromycin selection. (C) 2014 Elsevier Inc. All rights reserved.
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