A simple PCR method for rapid genotype analysis of the TH-MYCN transgenic mouse.
BACKGROUND
The TH-MYCN transgenic mouse is the most widely used murine model of human neuroblastoma, in which a human MYCN oncogene is targeted to neuroectodermal cells of developing mice under the influence of the rat tyrosine hydroxylase promoter. So far, homozygous transgenic mice have been identified by either Southern blot or quantitative real-time PCR.


RESULTS
To establish a simple and reliable genotyping method by conventional PCR, we confirmed the integration of the transgene in the TH-MYCN transgenic mouse by Southern blot and inverse PCR analyses. Our results showed that either five or six copies were found to be inserted in a head-to-tail tandem configuration at a single locus. The MYCN transgene/host DNA junction was sequenced and the integration site was identified at chromosome 18qE4. Finally, we succeeded in designing rapid, simple and reliable genotyping method by common PCR using primers flanking the integrated TH-MYCN transgene.


CONCLUSIONS
We established a simple and reliable genotyping PCR method for determining the integration site of the TH-MYCN transgene that enables all possible genotypes to be distinguished within several hours. TH-MYCN mice are excellent model for human neuroblastoma study, thus our results will largely be useful for facilitating the pace of neuroblastoma study, including in the study of the tumourigenic process, and in the development of therapies to treat patients suffering from neuroblastoma.
Haraguchi
Seiki
S
Laboratory of Embryonic and Developmental Genetics, Chiba Cancer Center Research Institute, Chiba, Japan.
Nakagawara
Akira
A
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D004195:Disease Models, Animal; D005260:Female; D005821:Genetic Techniques; D005838:Genotype; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009447:Neuroblastoma / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D019076:Transgenes; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
D016271:Proto-Oncogene Proteins c-myc; D014446:Tyrosine 3-Monooxygenase