Generation of an OCT3/4 reporter cynomolgus monkey ES cell line using CRISPR/Cas9

Publication date: Available online 15 April 2019Source: Stem Cell ResearchAuthor(s): Kenichi Kobayashi, Tomoyuki Tsukiyama, Masataka Nakaya, Susumu Kageyama, Keiji Tomita, Ryosuke Murai, Tetsuya Yoshida, Mitsuhiro Narita, Akihiro Kawauchi, Masatsugu EmaAbstractCynomolgus monkey ES (Cyn ES) cells can be generated in a similar manner as human ES cells. However, Cyn ES cells are difficult to maintain in an undifferentiated state by untrained researchers. For easier culture, we generated an OCT3/4-P2A tdTomato IRES ZeocinR Cyn ES cell line using CRISPR/Cas9 genome editing technology. The stop codon of the endogenous OCT3/4 locus was replaced with the P2A tdTomato IRES ZeocinR pA cassette by homologous recombination. This cell line enables us to isolate pluripotent stem cells and exclude differentiated cells by addition of zeocin, especially for culture without feeder cells.
Source: Stem Cell Research - Category: Stem Cells Source Type: research