Evaluation of tyrosinase minigene co-injection as a marker for genetic manipulations in transgenic mice.
The utility of tyrosinase minigene co-injection was evaluated as a visual marker for the generation and breeding of transgenic mice. In an evaluation of 39 transgenic founder animals and 44 transgenic lines five phenotypic patterns of pigmentation were consistently observed, including albino, dark, light, mottled and himalayan. In these studies co-injection of the tyrosinase minigene along with the transgene of interest (TOI) resulted in genomic integration of the two transgenes in 95% of the F0 generation. Co-segregation of transgenes occurred in 94% of doubly transgenic mice in the F1 generation, without dissociation in subsequent generations. All pigmented phenotypes proved useful for distinguishing homozygous from heterozygous F2 animals via backcross trials, while the light, mottled and himalayan phenotypes proved useful in visually discriminating between homozygous and heterozygous F2 animals. In addition, the light, mottled and himalayan phenotypes proved useful in determining segregation patterns of transgenes in the progeny of crosses between separate transgenic lines. Moreover, there appears to be a correlation between intensity of pigmentation and degree of expression of the co-injected TOI. These studies confirm that tyrosinase co-injection is a useful adjunct in transgenic mouse studies and can serve to reduce routine genetic validation of transgenic lines.
Institut de Recherches Cliniques de Montreal (IRCM), Faculty of Veterinary Medicine, University of Montreal, St.-Hyacinthe, Quebec, Canada.
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Reudelhuber
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D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D005091:Exons; D005260:Female; D015870:Gene Expression; D005819:Genetic Markers; D006200:Hair Color / Q000235:genetics*; D051379:Mice; D008813:Mice, Inbred ICR; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D005819:Genetic Markers; D014442:Monophenol Monooxygenase