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. 1998 Jul;4(7):468–479.

Subtyping analysis of Fanconi anemia by immunoblotting and retroviral gene transfer.

M Pulsipher 1, G M Kupfer 1, D Naf 1, A Suliman 1, J S Lee 1, P Jakobs 1, M Grompe 1, H Joenje 1, C Sieff 1, E Guinan 1, R Mulligan 1, A D D'Andrea 1
PMCID: PMC2230330  PMID: 9713825

Abstract

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.

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Selected References

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