Document Detail


Evaluating the Influence of Selection Markers on Obtaining Selected Pools and Stable Cell Lines in Human Cells.
MedLine Citation:
PMID:  23450727     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Selection markers are common genetic elements used in recombinant cell line development. While several selection systems exist for use in mammalian cell lines, no previous study has comprehensively evaluated their performance in the isolation of recombinant populations and cell lines. Here we examine four antibiotics, hygromycin, neomycin, puromycin, and Zeocin, and their corresponding selector genes, using a green fluorescent protein (GFP) as a reporter in two model cell lines, HT1080 and HEK293. We identify Zeocin as the best selection agent for cell line development in human cells. In comparison to the other selection systems, Zeocin is able to identify populations with higher fluorescence levels, which in turn leads to the isolation of better clonal populations and less false positives. Further, Zeocin-resistant populations exhibit better transgene stability in the absence of selection pressure compared to other selection agents. All isolated Zeocin-resistant clones, regardless of cell type, exhibited GFP expression. By comparison, only 79% of hygromycin-resistant, 47% of neomycin-resistant and 14% of puromycin-resistant clones expressed GFP. Based on these results, we would rank Zeocin > hygromycin ∼ puromycin > neomycin for cell line development in human cells. Furthermore, this study demonstrates that selection marker choice does impact cell line development.
Authors:
Amanda M Lanza; Do Soon Kim; Hal S Alper
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-3-1
Journal Detail:
Title:  Biotechnology journal     Volume:  -     ISSN:  1860-7314     ISO Abbreviation:  Biotechnol J     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-3-1     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101265833     Medline TA:  Biotechnol J     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Affiliation:
Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St. Stop C0400Austin, Texas 78712.
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