Array-based sequencing of filaggrin gene for comprehensive detection of disease-associated variants
| Autor: | Jianjun Liu, Rebecca L. Haines, Simon Denil, X.F. Colin C. Wong, Angeline S. L. Tay, Aileen Sandilands, Frances J.D. Smith, W.H. Irwin McLean, Mark B Y Tang, E. Birgitte Lane, Jia Nee Foo, John E.A. Common, Huijia Chen |
|---|---|
| Přispěvatelé: | Lee Kong Chian School of Medicine (LKCMedicine) |
| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine DNA Mutational Analysis Immunology Filaggrin Proteins Biology Genome Article Dermatitis Atopic law.invention 030207 dermatology & venereal diseases 03 medical and health sciences symbols.namesake 0302 clinical medicine Gene Frequency Intermediate Filament Proteins law Disease-associated Variants Humans Immunology and Allergy Coding region Medicine [Science] Exome Gene Alleles Exome sequencing Polymerase chain reaction Genetics Sanger sequencing Ichthyosis Filaggrin Gene 3. Good health 030104 developmental biology Mutation symbols Female Filaggrin |
| Zdroj: | The Journal of Allergy and Clinical Immunology |
| ISSN: | 0091-6749 |
| DOI: | 10.1016/j.jaci.2017.10.001 |
| Popis: | The filaggrin gene (FLG) is essential for skin differentiation and epidermal barrier formation. FLG loss-of-function (LoF) variants are associated with ichthyosis vulgaris and the major genetic risk factor for developing atopic dermatitis (AD).1, 2, 3 Genetic stratification of patients with AD according to FLG LoF risk is a common practice for both research and clinical studies; however, few studies comprehensively sequence the entire FLG coding region. Most studies that include FLG genotyping have screened for common predominant LoF variants to report allele frequencies after full Sanger sequencing of a smaller batch of test patient samples or previously published data. This strategy potentially results in underreporting of the genetic contribution especially in ethnicities where FLG LoF variants are highly diverse.4 Distinct LoF variants have been reported for most ethnicities studied to date. For example, 2 predominant sequence variants (p.R501X and c.2282del4) make up approximately 80% of the mutation burden in northern Europeans,5 whereas in East Asian ethnicities, a larger FLG LoF mutation spectrum is found with fewer predominating variants.6, 7 However, routinely Sanger sequencing the entire FLG coding region for large cohorts is not always feasible, although desirable as it is essential to correctly stratify patients. To address this, we developed a robust and cost-effective high-throughput PCR-based method for analyzing the entire coding region of FLG using Fluidigm microfluidics technology and next-generation sequencing (NGS). We have applied this method to fully resequence cohorts of Chinese, Malay, and Indian patients with AD from the Singaporean population. ASTAR (Agency for Sci., Tech. and Research, S’pore) Published version |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect