Weak D alleles in Japanese: a c.960G>A silent mutation in exon 7 of the RHD gene that affects D expression

Vox Sang. 2016 Feb;110(2):179-84. doi: 10.1111/vox.12322. Epub 2015 Sep 4.


Background and objectives: The molecular basis of the weak D phenotype has been investigated for many years, and more than 80 different alleles producing weak D phenotypes have been identified. Most alleles producing weak D phenotypes have a single missense mutation in exons corresponding to a transmembrane domain of the RhD polypeptide. We report here RHD alleles with single nucleotide mutations in Japanese accounting for weak expression of D antigen.

Methods: Seventy-five blood samples with a weak D phenotype were detected from 763 408 blood donors by standard serological methods. Forty-five of the 75 blood donors were available for RHD gene analysis by PCR and sequencing using genomic DNA and reticulocyte mRNA. Real-time PCR was performed to estimate the relative amounts of the RHD transcripts.

Results: We detected 16 different RHD alleles in the 45 individuals with weak D by nucleotide sequencing; 12 were newly identified. Thirty-two of the 45 individuals had an RHD allele with a single missense mutation, while the other 13 individuals had RHD with a c.960G>A silent mutation in exon 7. Red blood cells of these 13 individuals showed direct agglutination with anti-D at a strength of 3+ or less. Semi-quantitative analysis of the RHD transcripts by real-time PCR revealed that the cDNA samples with the c.960G>A mutation showed a significant increment of exon 7 skipping compared with the common RHD.

Conclusion: Reduced expression of D antigen is caused not only by missense mutation of the RHD gene, but also by silent mutation that may affect splicing.

Keywords: RHD gene; Rh blood group; exonic splicing regulator; silent mutation; weak D.

MeSH terms

  • Alleles*
  • Exons*
  • Humans
  • Mutation, Missense*
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Rh-Hr Blood-Group System / genetics*
  • Rh-Hr Blood-Group System / immunology
  • Rh-Hr Blood-Group System / metabolism
  • Silent Mutation*


  • RNA, Messenger
  • Rh-Hr Blood-Group System
  • Rho(D) antigen

Associated data

  • GENBANK/LC053435
  • GENBANK/LC053436
  • GENBANK/LC053437
  • GENBANK/LC053438
  • GENBANK/LC053439
  • GENBANK/LC053440
  • GENBANK/LC053441
  • GENBANK/LC053442
  • GENBANK/LC053443
  • GENBANK/LC053444
  • GENBANK/LC053445
  • GENBANK/LC053446
  • GENBANK/LC053447