Journal Article

Cell complementation using Genebridge 4 human:rodent hybrids for physical mapping of novel mitochondrial respiratory chain deficiency genes

Pascale de Lonlay, Claude Mugnier, Damien Sanlaville, Karine Chantrel-Groussard, Paule Bénit, Sophie Lebon, Dominique Chrétien, Noman Kadhom, Safa Saker, Gabor Gyapay, Serge Romana, Jean Weissenbach, Arnold Munnich, Pierre Rustin and Agnès Rötig

in Human Molecular Genetics

Volume 11, issue 26, pages 3273-3281
Published in print December 2002 | ISSN: 0964-6906
Published online December 2002 | e-ISSN: 1460-2083 | DOI: http://dx.doi.org/10.1093/hmg/11.26.3273
Cell complementation using Genebridge 4 human:rodent hybrids for physical mapping of novel mitochondrial respiratory chain deficiency genes

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The mapping and identification of respiratory chain deficiency genes is particularly tedious owing to the large number of genes encoding catalytic subunits and involved in respiratory chain (RC) assembly and maintenance. We have developed a functional complementation approach by: (i) growing the patient's fibroblasts in a highly selective medium; and (ii) transferring human chromosome fragments into RC-deficient fibroblasts by microcell-mediated transfer. In the absence of carbohydrates in the culture medium, the deficient cells rapidly disappeared unless they were rescued by a chromosome fragment carrying the disease gene. Microcells prepared from human:rodent Genebridge 4 panel of whole genome radiation hybrids were fused with fibroblast strains of two patients with complex II or I+IV deficiency and allowed to map the disease-causing genes to small intervals (4 and 12 Mb) on chromosomes 12p13 and 7p21, respectively. These intervals are similar to that obtained by genetic linkage analyses in large informative families. The recovery of normal RC enzyme activity in deficient skin fibroblasts supported the relevance of the transferred chromosome fragment in the disease. This approach makes the physical mapping of the disease genes feasible in some sporadic cases of RC deficiency.

Journal Article.  4641 words.  Illustrated.

Subjects: Genetics and Genomics

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