PURPOSE: Autosomal recessive retinitis pigmentosa (arRP) has recently been associated with mutations in a novel gene, EYS, which is a major gene for this disease. All published mutations so far are based on conventional PCR and are not adequate to identify midsized DNA rearrangements. This study was conducted to establish the prevalence of copy-number variations (CNVs) in the EYS gene in a cohort of arRP patients, including individuals in whom only one pathogenic change was detected by PCR-based sequencing. METHODS: A multiple ligation-dependent probe amplification (MLPA) was used for the molecular genetic analyses of CNVs by a novel EYS-specific kit. PCR-based direct sequencing was used in families where a pathogenic deletion or duplication was identified in one allele. Bioinformatics analyses was undertaken to study the effect of the mutations on protein structure and function. RESULTS: Six novel pathogenic CNVs were identified. Also, the presence of four midsized deletions was confirmed in patients previously identified. Midsized genomic rearrangements in EYS are disease causing in ∼4% of the families with no reported mutations and constitute the second pathogenic variation in ∼15% of cases where a mutation has been detected by direct sequencing. CONCLUSIONS: This is the first report of a systematic CNV screening of EYS gene in a cohort of arRP patients. Results suggest that midsized genomic rearrangements in EYS gene would be a common event in the appearance of RP phenotype. An efficient and cost-effective strategy validating a novel MLPA kit as a complementary diagnostic method for EYS pathogenic evaluation has been demonstrated.
PURPOSE:Autosomal recessive retinitis pigmentosa (arRP) has recently been associated with mutations in a novel gene, EYS, which is a major gene for this disease. All published mutations so far are based on conventional PCR and are not adequate to identify midsized DNA rearrangements. This study was conducted to establish the prevalence of copy-number variations (CNVs) in the EYS gene in a cohort of arRP patients, including individuals in whom only one pathogenic change was detected by PCR-based sequencing. METHODS: A multiple ligation-dependent probe amplification (MLPA) was used for the molecular genetic analyses of CNVs by a novel EYS-specific kit. PCR-based direct sequencing was used in families where a pathogenic deletion or duplication was identified in one allele. Bioinformatics analyses was undertaken to study the effect of the mutations on protein structure and function. RESULTS: Six novel pathogenic CNVs were identified. Also, the presence of four midsized deletions was confirmed in patients previously identified. Midsized genomic rearrangements in EYS are disease causing in ∼4% of the families with no reported mutations and constitute the second pathogenic variation in ∼15% of cases where a mutation has been detected by direct sequencing. CONCLUSIONS: This is the first report of a systematic CNV screening of EYS gene in a cohort of arRP patients. Results suggest that midsized genomic rearrangements in EYS gene would be a common event in the appearance of RP phenotype. An efficient and cost-effective strategy validating a novel MLPA kit as a complementary diagnostic method for EYS pathogenic evaluation has been demonstrated.
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Authors: Vera L Bonilha; Mary E Rayborn; Brent A Bell; Meghan J Marino; Gayle J Pauer; Craig D Beight; John Chiang; Elias I Traboulsi; Joe G Hollyfield; Stephanie A Hagstrom Journal: Graefes Arch Clin Exp Ophthalmol Date: 2014-12-11 Impact factor: 3.117
Authors: Tobias Eisenberger; Christine Neuhaus; Arif O Khan; Christian Decker; Markus N Preising; Christoph Friedburg; Anika Bieg; Martin Gliem; Peter Charbel Issa; Frank G Holz; Shahid M Baig; Yorck Hellenbroich; Alberto Galvez; Konrad Platzer; Bernd Wollnik; Nadja Laddach; Saeed Reza Ghaffari; Maryam Rafati; Elke Botzenhart; Sigrid Tinschert; Doris Börger; Axel Bohring; Julia Schreml; Stefani Körtge-Jung; Chayim Schell-Apacik; Khadijah Bakur; Jumana Y Al-Aama; Teresa Neuhann; Peter Herkenrath; Gudrun Nürnberg; Peter Nürnberg; John S Davis; Andreas Gal; Carsten Bergmann; Birgit Lorenz; Hanno J Bolz Journal: PLoS One Date: 2013-11-12 Impact factor: 3.240