FK506/Rapamycin-inducible Adenovirus Vector Construction Service

Rapamycin-inducible adenovirus vectors have been shown to regulate target gene transcription effectively in cell culture and in animals. Based on state-of-the-art adenovirus vectors technology platform and experienced team of experts, Creative Biolabs offers comprehensive rapamycin-inducible adenoviral vector construction services to achieve controlled expression of the gene.

Introduction of FK506/Rapamycin-inducible Expression System

Temporal and spatial control of gene expression is a fundamental tool for regulated protein expression for basic, pharmaceutical and clinical research. Several systems for controlling gene expression have been developed and the latest inducible gene expression system is the rapamycin-based dimerization. Rapamycin is a small molecule natural product that mediates the formation of heterodimers between the immunophilin FK506 binding protein (FKBP) and the lipid kinase homolog FRAP. In this system, the transcriptional transactivation domain is provided by the p65 subunit of the NFκB p65 protein, which is fused to the rapamycin-binding domain of FRAP. The DNA-binding domain, termed ZFHD1, is a composite zinc finger homeodomain chimeric protein with DNA recognition specificity fused to a series of three repeats of the FKBP. These two proteins dimerize in the presence of rapamycin to form a functional transactivator which binds an inducible promoter containing ZFHDI binding sites upstream of an hCMV or other minimal promoters to regulate the expression of target genes.

Rapamycin-regulated gene expression.Figure 1. Rapamycin-regulated gene expression. (Morsy, 1999)

Construction of FK506/Rapamycin-inducible Adenoviral Vectors

Rapamycin-inducible system has the potential to achieve tightly controlled gene regulation by expressing the transactivating and DNA-binding domains of a transcription factor as two separate proteins, each having a unique, highly specific rapamycin-binding domain, such that they dimerize to activate gene expression in the presence of rapamycin. This regulatable system can be integrated into adenoviral vectors to construct rapamycin-inducible adenoviral vectors to achieve controlled expression of the gene. In rapamycin-inducible adenoviral vectors, target genes and transcription factor components are delivered on isolated plasmids or on separate adenoviral vectors in vivo. Studies have shown that rapamycin-inducible adenoviral vectors are effective in controlling transgene expression in mice and non-human primates. One of the limitations of this approach is the growth inhibitory and immunosuppressive activity of rapamycin which is due to the inhibition of endogenous FRAP activity. This limitation can be overcome by developing nonimmunosuppressive analogs of rapamycin or modifying the FRAP sequence.

Service

With world-class expertise of viral vector technology, Creative Biolabs provides high-quality and one-stop rapamycin-inducible adenoviral vector design and construction services to meet the demand in basic research and preclinical applications. Compared to traditional adenoviral vectors, our rapamycin-inducible adenoviral vectors exhibit the following advantages:

  • The tightly regulated expression that allows for strict temporal control of adenovirus production.
  • The modified FRAP sequence allowed us to induce gene expression with nonimmunosuppressive rapamycin analogs.
  • This regulatory system enables long-term control of transgene expression in target cells.

In addition to the above services, Creative Biolabs also offers other regulated adenoviral vector construction services, including but not limited to the following:

No matter what stage, scope or scale of your adenovirus vector project, we can provide you with customized solutions to meet any of your needs. Please contact us if you have any questions or if you would like to place an order.

Reference

  1. Morsy, M.A.; et al. (1999). Helper-dependent adenoviral vectors as gene delivery vehicles. Gene Therapy and Molecular Biology. 3: 223-232.
For research use only. Not intended for any clinical use.