Heat-induced Adenovirus Vector Construction Service

Delivery of hsp70 promoter-driven transgene expression cassettes to tumor cells using adenoviral vector is a promising strategy for cancer therapy. Creative Biolabs has state-of-the-art facilities and experienced staff to assist the customers in all areas of adenoviral vector design and construction. With in-depth expertise in this field, we offer inducible adenovirus vector construction services with rapid turnaround time and lowest price.

Introduction of Heat-induced Gene Expression System

The heat shock response is a highly conserved process involving transient increases in the transcription of many heat shock genes when cells are subjected to heat stress. Heat shock gene expression is mediated through the interaction of an activated trimer of heat shock factor 1 (HSF-1) with a heat shock element (HSE) found in the promoters of heat-inducible genes. Studies have shown that some heat shock promoters, specifically, the hsp70 promoter, can activate thousands of times of gene expression in response to moderate hyperthermia. Incorporation of a heat shock gene promoter into a gene transfer vector has the potential to provide high levels of control of therapeutic gene expression in locally heated tumors. In the case of heat-inducible promoters, such as hsp70b, gene expression is typically targeted by local vector delivery, such as intratumoral adenovirus injection. Moreover, the addition of HSE to the hsp70 promoter improves inducibility and increases expression levels of the transgene.

Construction of Heat-induced Adenoviral Vectors

The hsp70b promoter can drive high-level transgene expression in the context of an adenoviral vector while maintaining relatively low levels of basal expression. By using the enhanced green fluorescent protein (EGFP) as a reporter gene, it was demonstrated that the expression of the heterologous gene controlled by the hsp70 promoter in the adenoviral vector could be elevated to 500-1000-fold over the background by moderate hyperthermia (39 °C to 43 °C) in tissue cultured cells. For example, under the control of heat shock promoter, a recombinant adenoviral vector containing EGFP was constructed to transduce exponential growth or plateau human prostate cancer cells (Dut-145 cells), which mediate high expression of EGFP (Figure 1). Furthermore, recombinant adenovirus vectors encoding the mouse IL-12 gene and the human TNF-ɑ gene, respectively, under the control of the hsp70b promoter have also been constructed and produced.

Fluorescent micrographs show heat-induced EGFP expression in Dut-145 cells infected at MOI 20 with adenovirus vectors, which contains the EGFP gene under control of the heat shock promoter.Figure 1. Fluorescent micrographs show heat-induced EGFP expression in Dut-145 cells infected at MOI 20 with adenovirus vectors, which contains the EGFP gene under control of the heat shock promoter. (Borrelli, 2001)

Application

The heat-induced gene expression system provides a promising approach to gene therapy that can be used to treat cancer and other diseases that require local or transient treatment of gene expression. The ability to localize and regulate expression from heat shock promoters in adenoviral vectors has proven to be particularly beneficial for many cancer applications, especially if the therapeutic products are highly toxic, such as proteotoxic or cytokines. Recently, heat-induced adenovirus vectors targeting of therapeutic genes such as IL-2, IL-12 and TNF-ɑ has shown good success in glioma and melanoma tumor models. Heat-induced adenoviral vector-mediated gene therapy shows some advantages over other therapies:

  • Heat-inducible gene transfer vectors may allow therapeutic genes to be targeted to locally heated tumors, thereby reducing nonspecific systemic gene expression.
  • The gene expression in vivo can be controlled and defined even when vectors are administered systemically.
  • No long-term side effects have been observed during decades of experimental cancer treatment.

Services

Creative Biolabs is a leading expert in adenoviral vector design and construction for gene delivery and gene therapy applications. Heat-induced adenoviral vectors have important application prospects in the fields of gene delivery and gene therapy. Based on our proprietary platform technology, we provide quality heat-induced adenoviral vector construction services to our global customers to meet their research and clinical needs.

In addition to the above services, other regulated adenoviral vector construction services are also available at Creative Biolabs, including:

If you are interested in our adenoviral vector construction services, please feel free to contact us.

Reference

  1. Borrelli, M.J.; et al. (2001). Heat-activated transgene expression from adenovirus vectors infected into human prostate cancer cells. Cancer Research. 61(3):1113-1121.
For research use only. Not intended for any clinical use.