Selection of intracellularly active ribozymes in mammalian cells
Hamada, M.; Kawasaki, H.; Kuwabara, T.; Warashina, M.; Taira, K.
Nucleic Acids Symposium Series 1999(42): 285-286
1999
ISSN/ISBN: 0261-3166 PMID: 10780491 Document Number: 501425
Ribozymes are expected to be useful as antiviral agents and powerful tools of functional analysis of unknown gene products in vivo. For use of ribozymes in vivo, they must be fully functional in the intracellular environment. Not all ribozymes selected in vitro would be expected to work in vivo, whereas ribozymes selected in the intracellular environment should retain their function in vivo. With the eventual aim of using ribozymes as antiviral agents or biological tools in mammalian cells, we then devised a novel selection system in mammalian cells of active ribozymes by targeting at a gene for the cyclin dependent kinase inhibitor (CDKI), p16INK4a. In this system, we found that p16INK4a-knockdown cells became malignant and they formed foci. In the mammalian system, we confirmed that the selected cells harbored the active ribozyme, indicating that our positive selection systems in vivo were operational.