Conventional subtraction library techniques or
DNA-transfection studies are standard techniques applied for identification and isolation of genes relevant in monogenetic diseases like
Fanconi anemia (FA). The differential display technique (
DDT) was developed to compare
mRNA expression between a mutant cell line and its syngeneic control and allows comparison of almost all
mRNA species within a short time. However, for identification of genes relevant in monogenetic diseases, no syngeneic cell model is available. In this report, we show that the use of nonsyngeneic diploid human fibroblasts does not increase the number of differentially displayed bands due to diversity of
untranslated regions.
cDNA bands with a length of up to 1000 bp were obtained and applied to
DDT. After screening of about 13000
cDNA bands, only 0.5% were found to be differentially expressed between FA and control cells. Finally, three mRNAs were cloned and verified in Northern blot experiments to be differentially expressed in FA fibroblasts. The low number of differentially displayed
cDNA bands in
DDT indicates the usefulness of this statistical, molecular approach for identification of multiple genes dysregulated in gene regulation cascades potentially relevant for cell cycle disturbances.