主催: Society for Reproduction and Development
Huntington disease (HD) is a fatal dominantly inherited neurodegenerative disease caused by the expansion of the polyCAG stretch in the gene coding ubiquitous huntingtin protein (htt). In order to facilitate studies of pathogenesis and therapeutics of HD we have generated transgenic miniature pig. F1 generation of HD transgenic miniature pigs is the offspring of F0 transgenic saw which was generated using microinjection of lentiviral vectors encoding N-truncated (548aa) human htt containing 145 polyQ repeats under the control of human htt promoter into the zygote. Genome analyses demonstrate the integration of transgene into the chromosome 1q24-q25 and the repetition of 124 glutamines in human htt. Assuming a presence of the two endogenous porcine htt alleles, we can announce that transgenic animals have integrated 1 insert of transgene in their genome. TR-FRET analysis of the F1 animals proved the active expression of human mutant htt in miniature pig´s cortical tissue. Western blot confirmed the expression of htt protein in the tissues from all three germ layers. F1 boars were able to produce F2 transgenic generations with the transgenic rate of approximately 50 % per birth. The first evident phenotype of Huntington disease has appeared in F1 transgenic boars at the age of 14 months. Reproductive parameters, number of spermatozoa per ejaculate, motility and in vitro penetration test, are gradually decreasing. The same phenotype has been discovered in F2 transgenic boars at the similar age. The preclinical stages in transgenic minipigs will provide a possibility to test in future all new pharmacological and molecular biology trials to find a disease modifying treatment of HD.
