One of the biggest challenges facing gene therapy is the development of vectors that direct the activity of therapeutic genes specifically to the sites of disease. To achieve this goal, the restriction of transgene transcription via synthetic promoters that are endowed with multiple specificities represents a particularly promising strategy. Towards this end, we have developed a generally applicable strategy (DCTF system) where a synthetic promoter is driven by an artificial heterodimeric transcription factor whose DNA-binding and transactivating subunits are expressed from two promoters with different selectivity. A crucial determinant of the DCTF system is the heterodimerization interface that should provide for a high affinity interaction without interference by endogenous proteins. Here, we describe such a dimerization system based on engineered Fos and Jun leucine zippers. We show the usefulness of this system for the combination of cell type-specific and cell cycle-regulated transcription and demonstrate its functionality in an in vivo setting.
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We are grateful to I Hart for the MeWo cell line, to N Favrot for the H322 cell line and to M Krause for oligonucleotide synthesis. We thank Simone Schmitt for performing the in vitro translation association assay. This work was supported by the Dr Mildred Scheel Stiftung.
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Jérôme, V., Müller, R. A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities. Gene Ther 8, 725–729 (2001). https://doi.org/10.1038/sj.gt.3301443
- cell cycle regulation
- DCTF system
- leucine zipper
- tissue-specific promoter
- transcriptional targeting
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