A series of plasmid vectors have been generated to allow the rapid construction of adenoviral vectors designed to express small RNA sequences. A truncated human U6 gene containing convenient restriction sites has been shown to be expressed at high levels following electroporation into a series of human cell lines. This gene was ligated into a promoterless adenoviral plasmid, and we have generated high titer virus by homologous recombination with adenoviral Addl327 DNA in 293 cells. Recombinant adenovirus containing a hammerhead ribozyme sequence targeted toward the Bcl-2 mRNA has been used to transduce a panel of human tumor cell lines. We have demonstrated high level expression of the recombinant U6 gene containing the ribozyme and reduction of Bcl-2 protein in transduced cells. These plasmids are suitable for the development of adenoviral vectors designed to express both ribozymes and antisense RNA in human cells.
Related information
Citations formats
Potter PM, McKenzie PP, Hussain N, Noonberg S, Morton CL, Harris LC (2000) Construction of adenovirus for high level expression of small RNAs in mammalian cells. Application to a Bcl-2 ribozyme Mol Biotechnol15: 105-14
Potter PM, McKenzie PP, Hussain N, Noonberg S, Morton CL, Harris LC (2000) Mol Biotechnol15: 105-14
