A Novel Missense Mutation in Van der Woude Syndrome: Usefulness of Fingernail DNA for Genetic Analysis

Abstract

Van der Woude syndrome (VWS) is an autosomal-dominant oral facial disorder. To find a gene mutation in a Japanese family using fingernail DNA samples, we performed this study. We hypothesized that a gene mutation in IRF6 might be involved in VWS, and that fingernail DNA samples may be valuable for detecting such mutations. Linkage and haplotype analyses of the family mapped the disease locus to the 1q32-q41 region. Mutation analysis with an improved extraction method for fingernail DNA detected a novel missense mutation (1046A>T, E349V) in exon 7 of IRF6 in all the affected members of the family. Since the E349V change may disturb the hydrophobic core and affect regulatory activity of IRF6, it is most likely that the mutation is causative for VWS in this family. Fingernail DNA is thus useful for linkage and mutation analyses, since the fingernail can be easily obtained non-invasively, sent through the mail, and stored for a long period. We emphasize here the usefulness of fingernail DNA for the genetic analysis of a disease.

Familial brain arteriovenous malformation maps to 5p13–q14, 15q11–q13 or 18p11: Linkage analysis with clipped fingernail DNA on high-density SNP array

“…Whole-blood samples cannot occasionally be available in some family members because of their far domicile. In such the case, fingernail DNA is useful, since clipped fingernails can be mailed in a usual way, and stored long at a room temperature, as indicated previously [22,27]. The present study is the first experience to adopt fingernail DNA to genome-wide …”

“…Genomic DNA was extracted from the fingernails using a buffer solution containing urea, DDT and proteinase K, as reported previously [22,23]. Briefly, clipped fingernails were once frozen in liquid nitrogen and crushed into fine powder using Multi-beads Shocker TM (Yasui Kikai, Osaka, Japan).…”

Genome-wide linkage analysis and mutation analysis of hereditary congenital blepharoptosis in a Japanese family

“…Double horizontal line depicts consanguineous marriage, and short bar above individual symbols indicates individuals examined clinically. Thick columns depict disease-associated haplotypes Linkage and haplotype analyses After obtaining written informed consent from each participant, DNA was extracted by conventional method from their whole blood, or using ISOHAIR TM (Nippon Gene, Tokyo, Japan) from their fingernail clippings, concentrated, and then purified by phenol-chloroform method (Matsuzawa et al 2006) For whole-genome scanning, we used the ABI Prism Linkage Mapping Set-MD10 (AppliedBiosystems, Foster City, CA, USA) that consists of 386 microsatellite markers from whole chromosomes with average distance of about 10 cM. Polymerase chain reaction (PCR) was performed in a 10-ll mixture containing 5 ng genomic DNA/0.25 U ExTaq DNA polymerase HS-version (TAKARA Bio Inc., Kyoto, Japan)/200 lM dNTP/0.3 lM primer/19 PCR buffer on the Dual 384-well GeneAmp PCR System 9700 Thermal Cycler (AppliedBiosystems).…”

Efficient DNA Extraction from Nail Clippings Using the Protease Solution from Cucumis melo

“…The collected nail clippings were cut into 2-3-mm squares using a new clean nail clipper. In the case of Frozen method [8], frozen nails in liquid nitrogen were pounded in a cold mortar to reduce them to a powder which was placed in 1.5 ml microtubes, washed three times in 70% EtOH, and then dried in air. Subsequently, 5 mg of nail clippings was added to a digestion buffer (80 ll) containing 1% SDS, 10 mM EDTA, and 10 mM CaCl 2 in 50 mM glycine buffer, pH 10.0, or 50 mM phosphate buffer, pH 7.5.…”

“…Some researchers have tried to solve this problem, and methods of improving the efficiency of DNA extraction have been established [6][7][8][9]. However, the yield has been varied.…”