Abstract |
Cyclin dependent kinases (cdks) play an important role in the pathogenesis of multiple neurodegenerative diseases. To explore the possibility of cdks-related gene therapy for neurodegenerative diseases, we packed recombinant adeno-associated virus (rAAV) encoding cdc2-siRNA. The expressing plasmid pAAV-MCS-EGFP-U6-cdc2-siRNA was constructed by using molecular biological techniques. The rAAV encoding cdc2-siRNA (rAAV-EGFP-U6-cdc2-siRNA) was packed by calcium phosphate mediated co-transfection of the plasmid pAAV-MCS-EGFP-U6-cdc2-siRNA, p-RC and p-Helper into AAV-293 cells. DNA sequencing proved the successful construction of U6-cdc2-siRNA in pAAV-MCS-EGFP. Seventy-two h after packaging, the expression of EGFP could be detected in AAV-293 cells. Western blotting revealed that cdc2 gene expression in AAV-293 cells was down-regulated markedly after transfection with rAAV-EGFP-U6-cdc2-siRNA, which evidenced the satisfactory silencing effect of this virus. It was concluded that the packaging of rAAV encoding cdc2-siRNA was successful. rAAV encoding cdc2-siRNA could silence cdc2 gene effectively, which might offer a novel means for the treatment of neurodegenerative diseases.
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Authors | Jiajun Wei, Min Zhang, Bitao Bu, Suming Zhang, Jinzhi Xu |
Journal | Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban
(J Huazhong Univ Sci Technolog Med Sci)
Vol. 28
Issue 6
Pg. 626-9
(Dec 2008)
ISSN: 1672-0733 [Print] China |
PMID | 19107353
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- RNA, Small Interfering
- enhanced green fluorescent protein
- Green Fluorescent Proteins
- CDC2 Protein Kinase
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Topics |
- Base Sequence
- CDC2 Protein Kinase
(genetics)
- Cell Line
- Dependovirus
(genetics, metabolism)
- Genetic Vectors
- Green Fluorescent Proteins
(genetics)
- Humans
- Molecular Sequence Data
- RNA, Small Interfering
(genetics)
- Recombination, Genetic
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