[Preimplantation diagnosis of delta F508 mutation of mucoviscidosis in transgenic mice].
OBJECTIVE
The aim of our study was to master preimplantation diagnosis in transgenic mice.


METHODS
Stage 4 or 8 cells preembryos were collected from C57BL6xCBAF1 mice, three days after mating with transgenic C57BL6xSJLF1 mice homozygous for the human gene delta F508 mutation of cystic fibrosis. A single blastomere was sampled by microbiopsy and the preembryos transferred in Swiss mice foster mothers. Molecular biology for delta F508 mutation on a single blastomere was performed with a double PCR technique on a Perkin Elmer Cetus 9600.


RESULTS
The success rate of biopsy on the 122 preembryos with 4 cells was 90.2%, and 93.8% on the 128 prembryos with 8 cells. The rate of in vitro hatching was 84.6% after biopsy on 52 preembryos with 4 cells (92.2% for controls without biopsy, p > 0.05), and 90.0% for 50 preembryos with 8 cells (94.4% for controls without biopsy, p > 0.05). The rate of birth after biopsy and transfer of 42 preembryos with 4 cells was 66.7% (72.5% for controls transferred without biopsy, p > 0.05) and 70.3% for 37 preembryos with 8 cells (71.9% for controls, p > 0.05). No difference was evidenced for births weight or organ weight at 3 weeks between mice born from biopsied embryos and controls. Thirty-two double PCR were performed for the diagnosis of the cystic fibrosis delta F508 mutation, 20 on a single blastomere obtained by microbiopsy and 12 for various negative controls, with 100% specificity and 100% sensitivity.


CONCLUSIONS
Microbiopsy of preembryos with 4 cells or 8 cells does not alter their viability or further development. The double PCR technique on a single blastomere for the cystic fibrosis delta F508 mutation is reliable with our current methods of investigation.
Ji
Y Z
YZ
Service de Gynécologie et Obstétrique, Hôpital Saint-Antoine, Paris.
Nicolas
J C
JC
Marechal
V
V
Desire
N
N
Pavirani
A
A
Milliez
J
J
D000818:Animals; D001707:Biopsy, Needle; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D047108:Embryonic Development; D005260:Female; D008722:Methods; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008967:Molecular Biology; D009154:Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011270:Pregnancy, Animal; D013997:Time Factors
Diagnostic préimplantatoire de la mutation delta F508 de la mucoviscidose chez la souris transgénique.