Abstract：
:Zinc-finger nucleases (ZFNs) are sequence-specific genome engineering tools with great potential for the development of gene therapies. The achievement of permanent cures through gene therapy requires targeting of stem cells but the effects and/or side effects of ZFN treatment on adult stem cell potency are largely unknown. Keratinocyte stem cells (KSCs) are attractive candidates for the development of gene therapies as their isolation, culture and grafting are well established. We derived KSCs from eGFP-transgenic mice and knocked out eGFP expression by disrupting the open reading frame with specific ZFNs in cell culture. EGFP-negative KSCs were then used as a model system to study the impact of ZFN treatment on stem cell potential. We achieved high gene disruption efficiencies with up to 18% eGFP-negative KSCs. As expected, ZFN cytotoxicity increased with rising ZFN concentrations. However, the ratio of correctly targeted KSCs among total treated cells was similar at different ZFN doses. Most importantly, our in vitro assays showed that ZFN-treated KSCs maintained their stem cell potential. They retained the capacity to both self-renew and form fully differentiated epidermal equivalents in culture. Moreover, they were able to form spherical aggregates in suspension culture, a characteristic hallmark shared with other stem cell types, and they expressed the in vivo KSC markers K15, NFATc1 and Sox9. Our data suggest that the stem cell potential of KSCs is not impaired by highly efficient ZFN treatment.
journal_name
Stem Cell Rev Repjournal_title
Stem cell reviews and reportsauthors
Höher T,Wallace L,Khan K,Cathomen T,Reichelt Jdoi
10.1007/s12015-011-9313-zsubject
Has Abstractpub_date
2012-06-01 00:00:00pages
426-34issue
2eissn
2629-3269issn
2629-3277journal_volume
8pub_type
杂志文章abstract：:During the process of development, neural crest cells migrate out from their niche between the newly formed ectoderm and the neural tube. Thereafter, they give rise not only to ectodermal cell types, but also to mesodermal cell types. Cell types with neural crest ancestry consequently comprise a number of specialized ...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2012-09-01 00:00:00
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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更新日期：2017-02-01 00:00:00
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2011-06-01 00:00:00
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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journal_title：Stem cell reviews and reports
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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更新日期：2012-03-01 00:00:00
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journal_title：Stem cell reviews and reports
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更新日期：2013-08-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
doi：10.1007/s12015-014-9521-4
更新日期：2014-10-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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journal_title：Stem cell reviews and reports
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pub_type： 杂志文章,评审
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更新日期：2019-06-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 已发布勘误
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更新日期：2019-02-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2020-08-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章
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更新日期：2010-12-01 00:00:00
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journal_title：Stem cell reviews and reports
pub_type： 杂志文章,评审
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更新日期：2019-04-01 00:00:00
abstract：:The original version of this article unfortunately contained a mistake. In the author group, the correct family name of Dr. Rebeca is "Blázquez" and the correct family name of Dr. Francisco Miguel is "Sánchez-Margallo." ...
journal_title：Stem cell reviews and reports
pub_type： 杂志文章,已发布勘误
doi：10.1007/s12015-020-09962-z
更新日期：2020-06-01 00:00:00
