Title: FANCA knockout in human embryonic stem cells causes a severe growth disadvantage
Authors: Vanuytsel, Kim ×
Cai, Qing
Nair, Nisha
Khurana, Satish
Shetty, Swati
Vermeesch, Joris
Ordovas Vidal, Laura
Verfaillie, Catherine #
Issue Date: Sep-2014
Publisher: Elsevier
Series Title: Stem Cell Research vol:13 issue:2 pages:240-50
Article number: 10.1016/j.scr.2014.07.005
Abstract: Fanconi anemia (FA) is an autosomal recessive disorder characterized by progressive bone marrow failure (BMF) during childhood, aside from numerous congenital abnormalities. FA mouse models have been generated; however, they do not fully mimic the hematopoietic phenotype. As there is mounting evidence that the hematopoietic impairment starts already in utero, a human pluripotent stem cell model would constitute a more appropriate system to investigate the mechanisms underlying BMF in FA and its developmental basis. Using zinc finger nuclease (ZFN) technology, we have created a knockout of FANCA in human embryonic stem cells (hESC). We introduced a selection cassette into exon 2 thereby disrupting the FANCA coding sequence and found that whereas mono-allelically targeted cells retain an unaltered proliferation potential, disruption of the second allele causes a severe growth disadvantage. As a result, heterogeneous cultures arise due to the presence of cells still carrying an unaffected FANCA allele, quickly outgrowing the knockout cells. When pure cultures of FANCA knockout hESC are pursued either through selection or single cell cloning, this rapidly results in growth arrest and such cultures cannot be maintained. These data highlight the importance of a functional FA pathway at the pluripotent stem cell stage.
ISSN: 1873-5061
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Interdepartemental Stem Cell Institute (-)
Stem Cell Biology and Embryology (+)
Department of Human Genetics - miscellaneous
× corresponding author
# (joint) last author

Files in This Item:
File Description Status SizeFormat
VAnuytsel et al 2014.pdf Published 1138KbAdobe PDFView/Open


All items in Lirias are protected by copyright, with all rights reserved.

© Web of science