Murakami K, Ohhira T, Oshiro E, Qi D, Oshimura M, Kugoh H
Department of Biomedical Science, Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Yonago, Japan.
Cytogenet Genome Res. 2009;125(1):19-25. doi: 10.1159/000207514. Epub 2009 Jul 14.
Xist non-coding RNA (ncRNA) is essential for X chromosome inactivation (XCI). Some genes can escape from XCI, but how this occurs is unknown. We developed a modified RNA tagging and recovery of associated proteins (TRAP) method to study the association between Xist RNA and its target genes. In mouse cells, Xist RNA was detected on the Uba1 gene, but not on Jarid1c and Utx genes, which escape from XCI. Using this technique we were able to show that the Xist RNA molecule is not present on active genes that escape from XCI, but is present on genes inactivated by XCI, suggesting that this method is a powerful tool for functional analysis of ncRNA.
Xist非编码RNA（ncRNA）对于X染色体失活（XCI）至关重要。一些基因能够逃避XCI，但具体机制尚不清楚。我们开发了一种改良的RNA标记及相关蛋白回收（TRAP）方法，以研究Xist RNA与其靶基因之间的关联。在小鼠细胞中，Uba1基因上可检测到Xist RNA，但逃避XCI的Jarid1c和Utx基因上则未检测到。利用该技术我们能够证明，Xist RNA分子不存在于逃避XCI的活性基因上，而是存在于因XCI而失活的基因上，这表明该方法是用于ncRNA功能分析的有力工具。
