Abstract
Germ line mutations in genes involved in hereditary cancer syndromes, such as BRCA1 and BRCA2 in breast cancer and MSH2, MSH6, MLH1, and PSM2 in hereditary nonpolyposis colorectal cancer (HNPCC, more recently indicated as Lynch syndrome), confer a high risk to develop cancer. Mutation analysis in these genes has resulted in the identification of a large number of sequence variants, of which mutations causing frame shifts and nonsense codons are considered undoubtedly to be pathogenic. Many variants, however, cannot be classified as either disease-causing mutations or neutral variants and are therefore called unclassified variants (UVs). A subset of these variants may have an effect on RNA splicing. Appropriate RNA analysis will enable the characterization of the exact molecular nature of this effect and hence, is essential to determine the clinical relevance of the genomic variant. This chapter describes the design and implementation of RNA analysis as an indispensible tool in today’s clinical diagnostic setting.
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Acknowledgments
We acknowledge support from the Dutch Cancer Society Grant number 2001-2471 (M.V.).
We thank our Dutch and Belgium colleagues of the DNA diagnostic laboratories for the discussions on the methodologies for RNA analysis and J.T. Wijnen, Ph.D. (Department of Clinical Genetics, Leiden University Medical Center, Leiden), A. van den Ouweland, Ph.D. (of Clinical Genetics, Erasmus Medical Center, Rotterdam) and R. Blok, Ph.D. (Dept. of Genetics and Cell Biology, Maastricht University Medical Center, Maastricht), The Netherlands, for critical reading of the manuscript.
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Vreeswijk, M.P.G., van der Klift, H.M. (2012). Analysis and Interpretation of RNA Splicing Alterations in Genes Involved in Genetic Disorders. In: Aartsma-Rus, A. (eds) Exon Skipping. Methods in Molecular Biology, vol 867. Humana Press. https://doi.org/10.1007/978-1-61779-767-5_4
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DOI: https://doi.org/10.1007/978-1-61779-767-5_4
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