Please use this identifier to cite or link to this item: https://doi.org/10.7150/ijbs.6507
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
Nanos3
Pluripotency
Issue Date: 2013
Source: 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. https://doi.org/10.7150/ijbs.6507
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
URI: http://scholarbank.nus.edu.sg/handle/10635/101179
ISSN: 14492288
DOI: 10.7150/ijbs.6507
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