Abstract
mRNA is an important tool to study the effects of particular mutations on the mode of splicing and transcripts. However, it is often difficult to isolate mRNA because the organ or tissue in which the gene is expressed cannot be sampled. We previously identified two probable splicing mutations (c.6485+5G>A and c.8559-2A>G) during the mutation analysis of USH2A in Japanese Usher syndrome (USH) type 2 patients, but we could not observe their effects on splicing because the gene is expressed in only a few tissues/organs, and is not expressed in peripheral lymphocytes. In this study, we used hair roots as a source of mRNA of USH-causing genes, and successfully detected the expression of seven, except USH1C and CLRN1, of the nine USH-causing genes. We used RNA extracted from the hair roots of a patient who has both c.6485+5G>A and c.8559-2A>G mutations in USH2A in a compound heterozygous state to observe the effects of these mutations on transcripts. Reverse-transcription PCR analysis revealed that c.6485+5G>A and c.8559-2A>G inactivated splice donor and splice acceptor sites, respectively, and caused skipping of exons. Thus, RNA extracted from hair roots is a potential powerful and convenient tool for the mutation analysis of USH-causing genes.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Nakanishi, H., Ohtsubo, M., Iwasaki, S., Hotta, Y., Mizuta, K., Mineta, Y. et al. Identification of 11 novel mutations in USH2A among Japanese patients with Usher syndrome type 2. Clin. Genet. 76, 383–391 (2009).
van Wijk, E., Pennings, R. J., te Brinke, H., Claassen, A., Yntema, H. G., Hoefsloot, L H. et al. Identification of 51 novel exons of the Usher syndrome type 2A (USH2A) gene that encode multiple conserved functional domains and that are mutated in patients with Usher syndrome type II. Am. J. Hum. Genet. 74, 738–744 (2004).
Ebermann, I., Scholl, H. P., Charbel Issa, P., Becirovic, E., Lamprecht, J., Jurklies, B. et al. A novel gene for Usher syndrome type 2: mutations in the long isoform of whirlin are associated with retinitis pigmentosa and sensorineural hearing loss. Hum. Genet. 121, 203–211 (2007).
Wolfrum, U., Liu, X., Schmitt, A., Udovichenko, I. P. & Williams, D. S. Myosin VIIa as a common component of cilia and microvilli. Cell Motil. Cytoskeleton. 40, 261–271 (1998).
Nakai, K. & Sakamoto, H. Construction of a novel database containing aberrant splicing mutations of mammalian genes. Gene. 141, 171–177 (1994).
Cartegni, L., Chew, S. L. & Krainer, A. R. Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat. Rev. Genet. 3, 285–298 (2002).
Acknowledgements
This work was supported by research grants from the Ministry of Health, Labour and Welfare (Acute Profound Deafness Research Committee), and from the Ministry of Education, Culture, Sports, Science and Technology (Young Scientists Grant B-20791189) in Japan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nakanishi, H., Ohtsubo, M., Iwasaki, S. et al. Hair roots as an mRNA source for mutation analysis of Usher syndrome-causing genes. J Hum Genet 55, 701–703 (2010). https://doi.org/10.1038/jhg.2010.83
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/jhg.2010.83
Keywords
This article is cited by
-
A novel USH2A variant in a patient with hearing loss and prenatal diagnosis of a familial fetus: a case report
BMC Medical Genomics (2021)
-
The first USH2A mutation analysis of Japanese autosomal recessive retinitis pigmentosa patients: a totally different mutation profile with the lack of frequent mutations found in Caucasian patients
Journal of Human Genetics (2014)
-
Screening for duplications, deletions and a common intronic mutation detects 35% of second mutations in patients with USH2A monoallelic mutations on Sanger sequencing
Orphanet Journal of Rare Diseases (2013)
-
Novel USH2A mutations in Japanese Usher syndrome type 2 patients: marked differences in the mutation spectrum between the Japanese and other populations
Journal of Human Genetics (2011)
-
Mutation analysis of the MYO7A and CDH23 genes in Japanese patients with Usher syndrome type 1
Journal of Human Genetics (2010)
