Abstract
Objective
To develop a mouse artificial chromosome (MAC) carrying the mouse Xist gene (X-inactive specific transcript; Xist-MAC) as a systematic in vitro approach for investigating Xist RNA-mediated chromosome inactivation.
Results
Ectopic expression of the Xist gene in CHO cells led to the accumulation of Xist RNA in cis on the MAC. In addition, the introduction of Xist-MAC to embryonic stem cells from male mice via microcell-mediated chromosome transfer resulted in the accumulation of Xist RNA in cis on the MAC. Chromosomal inactivation was observed in the differentiated state. Moreover, this phenomenon was accompanied by the epigenetic modification of H3K27 trimethylation.
Conclusions
We successfully generated a novel chromosome inactivation model, Xist-MAC, which will provide a valuable tool for the screening and functional analysis of X chromosome inactivation-related genes and proteins.
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Acknowledgements
We are grateful to Tatsuya Ohhata for generously providing us with the J1:rtTA ES cells, to Sayaka Aoki and Rikako Yagura for helping with our experiments, and to Dr. Yasuhiro Kazuki and Dr. Mitsuo Oshimura. This research was partly performed at the Tottori Bio Frontier managed by Tottori prefecture. We also thank Susan Furness, PhD, from Edanz Group (https://en-author-services.edanzgroup.com/) for editing a draft of this manuscript.
Funding
This work was supported by the Japan Society for the Promotion of Science [Grant Nos. JP17J03813 and JP25640114, in part].
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Inaoka, D., Sunamura, N., Ohira, T. et al. A novel Xist RNA-mediated chromosome inactivation model using a mouse artificial chromosome. Biotechnol Lett 42, 697–705 (2020). https://doi.org/10.1007/s10529-020-02826-z
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DOI: https://doi.org/10.1007/s10529-020-02826-z