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Thorough in silico and in vitro cDNA analysis of 21 putative BRCA1 and BRCA2 splice variants and a complex tandem duplication in BRCA2 allowing the identification of activated cryptic splice donor sites in BRCA2 exon 11
Author(s) -
Baert Annelot,
Machackova Eva,
Coene Ilse,
Cremin Carol,
Turner Kristin,
PortigalTodd Cheryl,
Asrat Marie Jill,
Nuk Jennifer,
Mindlin Allison,
Young Sean,
MacMillan Andree,
Maerken Tom,
Trbusek Martin,
McKin Wendy,
Wood Marie E.,
Foulkes William D.,
Santamariña Marta,
la Hoya Miguel,
Foretova Lenka,
Poppe Bruce,
Vral Anne,
Rosseel Toon,
Leeneer Kim,
Vega Ana,
Claes Kathleen B. M.
Publication year - 2018
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.23390
Subject(s) - biology , in silico , exon , rna splicing , genetics , splice , tandem exon duplication , computational biology , breakpoint , intron , exon skipping , complementary dna , alternative splicing , gene , rna , chromosomal translocation
For 21 putative BRCA1 and BRCA2 splice site variants, the concordance between mRNA analysis and predictions by in silico programs was evaluated. Aberrant splicing was confirmed for 12 alterations. In silico prediction tools were helpful to determine for which variants cDNA analysis is warranted, however, predictions for variants in the Cartegni consensus region but outside the canonical sites, were less reliable. Learning algorithms like Adaboost and Random Forest outperformed the classical tools. Further validations are warranted prior to implementation of these novel tools in clinical settings. Additionally, we report here for the first time activated cryptic donor sites in the large exon 11 of BRCA2 by evaluating the effect at the cDNA level of a novel tandem duplication (5′ breakpoint in intron 4; 3′ breakpoint in exon 11) and of a variant disrupting the splice donor site of exon 11 (c.6841+1G > C). Additional sites were predicted, but not activated. These sites warrant further research to increase our knowledge on cis and trans acting factors involved in the conservation of correct transcription of this large exon. This may contribute to adequate design of ASOs (antisense oligonucleotides), an emerging therapy to render cancer cells sensitive to PARP inhibitor and platinum therapies.