Rapid fluorescence in situ hybridization on interphasic nuclei to discriminate between homozygous and heterozygous transgenic mice.
Homozygous and heterozygous transgenic mice of the Tg152 line overexpressing the human copper/zinc superoxide dismutase (hSOD-1) were rapidly differentiated by fluorescence in situ hybridization (FISH) using interphase lymphocyte nuclei. We have devised a simple and fast method for preparing interphase nuclei with very small quantities of whole mouse blood, avoiding several steps of the classical FISH technique. Lymphocyte separation and cell culture were not required. This technique provides an excellent tool for the unambiguous detection of homozygous and heterozygous transgenic mice in a litter. It can be used to check young animals since 2 microliters of whole blood is sufficient. We also show that in this transgenic line numerous copies of the hSOD-1 transgene are integrated at a single autosomal locus, in tandem head-to-tail organization.
Paris
D
D
CNRS URA 1335, Faculté de Médecine Necker-Enfants Malades, Paris, France.
Toyama
K
K
Mégarbané
A
A
Casanova
P M
PM
Sinet
P M
PM
London
J
J
D000818:Animals; D015139:Blotting, Southern; D001947:Breeding; D002467:Cell Nucleus / Q000235:genetics*; D002875:Chromosomes; D005260:Female; D005787:Gene Frequency; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000737:chemistry / Q000201:enzymology; D008677:Metaphase; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
D013482:Superoxide Dismutase