Transgenic mice generated by pronuclear injection of a yeast artificial chromosome.
Transgenic mice have become invaluable for analysing gene function and regulation in vivo. However, the size of constructs injected has been limited by the cloning capacity of conventional vectors, a constraint that could be overcome with yeast artificial chromosomes (YACs). We investigated the feasibility of making transgenic mice with YACs by pronuclear injection of a small YAC carrying a gene encoding tyrosinase. Use of a vector with a conditional centromere allowed fifteenfold amplification of the YAC in yeast and its recovery in high yield. The albino phenotype of the recipient mice was rescued demonstrating the correct expression of the tyrosine gene from the construct. Furthermore, the telomeric sequences added by the yeast integrated into the mouse genome and did not reduce efficiency of integration. Using this technique future experiments with longer YACs will allow the expression of gene complexes such as Hox and the globin gene clusters to be analysed in transgenic animals.
Schedl
A
A
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Beermann
F
F
Thies
E
E
Montoliu
L
L
Kelsey
G
G
Schütz
G
G
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D016678:Genome; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D008845:Microinjections; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000378:metabolism; D015053:Zygote / Q000378:metabolism*
D005819:Genetic Markers; D014442:Monophenol Monooxygenase