Subtyping analysis of Fanconi anemia by immunoblotting and retroviral gene transfer

Mol Med. 1998 Jul;4(7):468-79.


Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with at least eight complementation groups (A-H). Two of the FA genes (FAA and FAC) have been cloned, and mutations in these genes account for approximately 80% of FA patients. Subtyping of FA patients is an important first step toward identifying candidates for FA gene therapy. In the current study, we analyzed a reference group of 26 FA patients of known subtype. Most of the patients (18/26) were confirmed as either type A or type C by immunoblot analysis with anti-FAA and anti-FAC antisera. In order to resolve the subtype of the remaining patients, we generated retroviral constructs expressing FAA and FAC for transduction of FA cell lines (pMMP-FAA and pMMP-FAC). The pMMP-FAA construct specifically complemented the abnormal phenotype of cell lines from FA-A patients, while pMMP-FAC complemented FA-C cells. In summary, the combination of immunoblot analysis and retroviral-mediated phenotypic correction of FA cells allows a rapid method of FA subtyping.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Western / methods
  • Cell Cycle / drug effects
  • Cell Cycle Proteins*
  • Cell Line, Transformed
  • Cells, Cultured
  • DNA-Binding Proteins*
  • Fanconi Anemia / classification*
  • Fanconi Anemia / diagnosis
  • Fanconi Anemia / genetics*
  • Fanconi Anemia Complementation Group Proteins
  • Fibroblasts / drug effects
  • Gene Transfer Techniques*
  • Genetic Complementation Test*
  • Genetic Therapy
  • Humans
  • Immune Sera
  • Lymphocytes / drug effects
  • Mitomycin / pharmacology
  • Molecular Sequence Data
  • Nuclear Proteins*
  • Proteins / analysis*
  • Proteins / genetics
  • Retroviridae / genetics


  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Fanconi Anemia Complementation Group Proteins
  • Immune Sera
  • Nuclear Proteins
  • Proteins
  • Mitomycin