Abstract
As a result of X chromosome inactivation, females are mosaic for cell lineages in which either the paternal or the maternal X chromosome is active, and, if inactivation were random, each lineage should be present at approximately the same frequency. Detection of instances of non-random X inactivation can be important both clinically and for the study of X chromosome inactivation. Identification of a single-base polymorphism in an expressed region of the human XIST gene has permitted the development of a direct PCR-based assay for randomness of X inactivation. Oligonucleotide primers were designed, incorporating the variant base, and conditions established that allowed allele-specific PCR amplification. As the XIST gene is expressed only from the inactive X chromosome, differential amplification of the alleles in cDNA from heterozygotes can be used as an indicator of non-random inactivation. Using this assay, non-random X chromosome inactivation has been demonstrated in chromosomally abnormal cell lines and in lymphocytes from heterozygous, normal females. Virtually complete non-random X inactivation was also shown in a mother and her daughter, suggesting the existence of some familial factor affecting X chromosome inactivation.
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Acknowledgements
We would like to thank Karen Gustashaw and Dr. Denise Saker for their assistance in obtaining the blood samples and preparing the DNA and RNA. We thank Dr. Alasdair Hunter (Ottawa, Canada) for providing samples from the family with X-linked ichthyosis. This work was supported by research grant GM45441 from the National Institutes of Health to H.F.W.
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Rupert, J.L., Brown, C.J. & Willard, H.F. Direct Detection of Non-Random X Chromosome Inactivation by Use of a Transcribed Polymorphism in the XIST Gene. Eur J Hum Genet 3, 333–343 (1995). https://doi.org/10.1159/000472322
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DOI: https://doi.org/10.1159/000472322
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