International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)

Table of Contents * Complex Genetics * Developmental Genetics * Gene Annotation * Gene Discovery * Genome Sequencing * Functional Genomics * Mutagenesis * Presentations * Verne Chapman Memorial Lecture

A GENE-DRIVEN APPROACH TO THE IDENTIFICATION OF ENU MUTANTS IN THE MOUSE CONNEXIN 26 GENE

Emma Coghill
MRC Harwell
MRC Mammalian Genetics Unit & UK Mouse Genome Centre,
Harwell,
Oxon, OX11 ORD

Co-Authors: 1)Hugill A, 2)Hunter AJ, 1)Cox R, 1)Brown SDM
Institutions: 1)MRC Mammalian Genetics Unit and UK Mouse Genome Centre, 2)GlaxoSmithKline

A gene-driven approach to the identification of enu mutants in the mouse connexin 26 gene

The establishment of parallel DNA and sperm archives from ENU mutagenised mice represents a powerful approach to gene-driven mutagenesis, allowing for rapid DNA screening and swift mutant recovery. A spectrum of single base changes could be recovered, potentially uncovering the full array of functional changes including hypomorphs, antimorphs and neomorphs. We provide proof of principle of this gene-driven approach, reporting the identification of a nonsense mutation in the connexin 26 (Gjb2) deafness gene. GJB2 gene mutations are the most common cause of human non-syndromic deafness. A mouse Gjb2 gene knock-out is embryonic lethal, therefore, no mouse model for this form of deafness is currently available. The gene-driven approach was used in an attempt to recover new alleles at the mouse Gjb2 locus and investigate their phenotype. Screening of 500 archive DNAs, identified one mouse carrying a stop mutation at codon 119, which was recovered by IVF and the Gjb2 stop mutation confirmed. Heterozygous mice showed normal hearing and intercrosses produced no homozygous progeny, demonstrating embryonic lethality. We have begun to apply a high-throughput gene-driven approach for the recovery of mouse mutants in a variety of cochlear-expressed genes, including a search for further alleles of the Gjb2 gene.