| 摘要 |
The invention relates to the field of molecular biology, virology, and gene therapy where various vectors, particularly viral and retroviral vectors, might be used for gene transfer either into the cultivated cells in vitro or into living organisms. Specifically, the invention provides solution to the problem of silenced gene expression and methylation of the vector DNA in eukaryotic cells, particularly in mammalian cells. This solution consists in the use of a specific regulatory gene element containing a promoter/enhancer based on the retroviral long terminal repeat (LTR) modified by insertion of at least one internal element (IE) from the CpG island. The LTR modified in this way ensures strong and long-term expression of the integrated vector together with the transduced gene of interest (transgene) and, at he same time, its efficient protection from transcriptional silencing and gradual methylation, even in human cells. Particularly, the invention discloses a regulatory gene element comprising the enhancer/promoter protected from DNA methylation and from silencing of gene expression, wherein said element comprises a modified retroviral sequence LTR with at least one IE sequence inserted in any orientation into any position, wherein the IE sequence comprises SEQUENCE ID No. 2. or any equivalent sequence that fulfils the same function. |
