Members of cdc2 family of protein kinases are known for their pivotal role in the regulation of the eukaryotic cell cycle. The potential roles of neuronal specific cdc2-like kinases in stabilizing neurofilament skeleton and in axonal morphogenesis through tau by phosphorylation are not well delineated. The main objectives of this project are: (1) to disrupt the genomic locus of neuronal cdc2-like kinase (cdk5) in embryonic stem cells, and then use the targeted cells to generate mouse models to study in vivo function of these kinases; and (2) to overexpress cdk5 in neuronal cell lines and in vivo in mice. Toward achieving these objectives, we have isolated 8 genomic clones of cdk5 from the 129/svj library. These isogeneic clones have been partially characterized to define intron/exon boundaries and exonic sequences. A gene-targeting construct has been engineered with a deletion of exons 3 through 5 and insertion of neomycin resistance gene at the site of deletion. At the 3' end of the construct, the herpes virus thymidine kinase gene is included for positive/negative selection using G418 and gancyclovir. We have also isolated a murine cDNA clone using the RT-PCR technique, and have obtained complete nucleic acid sequence data. Gene targeting experiments in ES are currently in progress to obtain targeted clones.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Intramural Research (Z01)
Project #
1Z01NS002893-01
Application #
3760370
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
City
State
Country
United States
Zip Code