title: Analysis and interpretation of RNA splicing alterations in genes involved in genetic disorders.
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.
authors.affiliation: Department of Human Genetics, Center for Human and Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands. vreeswijk@lumc.nl
authors.forename: Maaike P G
authors.initials: MP
authors.lastname: Vreeswijk
authors.forename: Heleen M
authors.initials: HM
authors.lastname: van der Klift
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005347:Fibroblasts / Q000378:metabolism; D020022:Genetic Predisposition to Disease / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D018095:Germ-Line Mutation; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D009154:Mutation; D009369:Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification; D012326:RNA Splicing
chemical_list: D018076:DNA, Complementary; D012313:RNA; D004247:DNA