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Proof-of-concept fo...
Proof-of-concept for CRISPR/Cas9 gene editing in human preadipocytes : Deletion of FKBP5 and PPARG and effects on adipocyte differentiation and metabolism
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- Kamble, Prasad G. (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Hetty, Susanne, 1979- (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Vranic, Milica (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Almby, Kristina (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Castillejo-Lopez, Casimiro (author)
- Uppsala universitet,Medicinsk genetik och genomik,Science for Life Laboratory, SciLifeLab
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- Abalo, Xesús M., 1976- (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Pereira, Maria J., 1981- (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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- Eriksson, Jan W. (author)
- Uppsala universitet,Klinisk diabetologi och metabolism
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(creator_code:org_t)
- 2020-06-29
- 2020
- English.
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In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 10:1
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Abstract
Subject headings
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- CRISPR/Cas9 has revolutionized the genome-editing field. So far, successful application in human adipose tissue has not been convincingly shown. We present a method for gene knockout using electroporation in preadipocytes from human adipose tissue that achieved at least 90% efficiency without any need for selection of edited cells or clonal isolation. We knocked out the FKBP5 and PPARG genes in preadipocytes and studied the resulting phenotypes. PPARG knockout prevented differentiation into adipocytes. Conversely, deletion of FKBP51, the protein coded by the FKBP5 gene, did not affect adipogenesis. Instead, it markedly modulated glucocorticoid effects on adipocyte glucose metabolism and, furthermore, we show some evidence of altered transcriptional activity of glucocorticoid receptors. This has potential implications for the development of insulin resistance and type 2 diabetes. The reported method is simple, easy to adapt, and enables the use of human primary preadipocytes instead of animal adipose cell models to assess the role of key genes and their products in adipose tissue development, metabolism and pathobiology.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Endokrinologi och diabetes (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Endocrinology and Diabetes (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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