Silencing of the Pink1 gene expression by conditional RNAi does not induce dopaminergic neuron death in mice.
Transgenic RNAi, an alternative to the gene knockout approach, can induce hypomorphic phenotypes that resemble those of the gene knockout in mice. Conditional transgenic RNAi is an attractive choice of method for reverse genetics in vivo because it can achieve temporal and spatial silencing of targeted genes. Pol III promoters such as U6 are widely used to drive the expression of RNAi transgenes in animals. Tested in transgenic mice, a Cre-loxP inducible U6 promoter drove the broad expression of an shRNA against the Pink1 gene whose loss-of-functional mutations cause one form of familial Parkinson's disease. The expression of the shRNA was tightly regulated and, when induced, silenced the Pink1 gene product by more than 95% in mouse brain. However, these mice did not develop dopaminergic neurodegeneration, suggesting that silencing of the Pink1 gene expression from embryo in mice is insufficient to cause similar biochemical or morphological changes that are observed in Parkinson's disease. The results demonstrate that silencing of the PINK1 gene does not induce a reliable mouse model for Parkinson's disease, but that technically the inducible U6 promoter is useful for conditional RNAi in vivo.
Zhou
Hongxia
H
Department of Pathology, Anatomy & Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Falkenburger
Björn H
BH
Schulz
Jörg B
JB
Tieu
Kim
K
Xu
Zuoshang
Z
Xia
Xu Gang
XG
D000818:Animals; D002460:Cell Line; D003433:Crosses, Genetic; D018076:DNA, Complementary / Q000235:genetics; D004298:Dopamine / Q000032:analysis; D005786:Gene Expression Regulation; D006801:Humans; D007668:Kidney; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology*; D010300:Parkinson Disease / Q000235:genetics; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D012342:RNA, Small Nuclear / Q000235:genetics; D013045:Species Specificity; D016385:TATA Box; D014162:Transfection; D000099202:PTEN-Induced Putative Kinase
D018076:DNA, Complementary; D004298:Dopamine; D011494:Protein Kinases; D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; D000099202:PTEN-Induced Putative Kinase; C405486:U6 small nuclear RNA