Please use this identifier to cite or link to this item:
Title: Nanos3 gene targeting in medaka ES cells
Authors: Guan, G.
Yan, Y. 
Chen, T. 
Yi, M.
Ni, H.
Naruse, K.
Nagahama, Y.
Hong, Y. 
Keywords: ES
Gene targeting
Homologous recombination
Issue Date: 2013
Citation: Guan, G., Yan, Y., Chen, T., Yi, M., Ni, H., Naruse, K., Nagahama, Y., Hong, Y. (2013). Nanos3 gene targeting in medaka ES cells. International Journal of Biological Sciences 9 (5) : 444-454. ScholarBank@NUS Repository.
Abstract: Gene targeting (GT) by homologous recombination offers the best precision for genome editing in mice. nanos3 is a highly conserved gene and encodes a zinc-finger RNA binding protein essential for germ stem cell maintenance in Drosophila, zebrafish and mouse. Here we report nanos3 GT in embryonic stem (ES) cells of the fish medaka as a lower vertebrate model organism. A vector was designed for GT via homologous recombination on the basis of positive-negative selection (PNS). The ES cell line MES1 after gene transfer and PNS produced 56 colonies that were expanded into ES cell sublines. Nine sublines were GT-positive by PCR genotyping, 4 of which were homologous recombinants as revealed by Southern blot. We show that one of the 4, A15, contains a precisely targeted nanos3 allele without any random events, demonstrating the GT feasibility in medaka ES cells. Importantly, A15 retained all features of undifferentiated ES cells, including stable self-renewal, an undifferentiated phenotype, pluripotency gene expression and differentiation during chimeric embryogenesis. These results provide first evidence that the GT procedure and genuine GT on a chromosomal locus such as nanos3 do not compromise pluripotency in ES cells of a lower vertebrate. © Ivyspring International Publisher.
Source Title: International Journal of Biological Sciences
ISSN: 14492288
DOI: 10.7150/ijbs.6507
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.


checked on Apr 6, 2020


checked on Mar 26, 2020

Page view(s)

checked on Mar 28, 2020

Google ScholarTM



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.