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Updated: Jul 17, 2026
Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
Published on: April 27, 2018
Novel system uses probasin-based promoter, transcriptional silencers and amplification loop to induce high-level
Jan Woraratanadharm1, Semyon Rubinchik, Hong Yu
1Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, South Carolina, USA. jworara@genphar.com <jworara@genphar.com>
Background:
Despite several effective treatment options available for prostate cancer, it remains the second leading cause of cancer death in American men. Thus, there is a great need for new treatments to improve outcomes. One such strategy is to eliminate cancer through the expression of cytotoxic genes specifically in prostate cells by gene therapy vectored delivery. To prevent systemic toxicity, tissue- and/or cancer-specific gene expression is required. However, the use of tissue- or cancer-specific promoters to target transgene expression has been hampered by their weak activity.
Results:
To address this issue, we have developed a regulation strategy that includes feedback amplification of gene expression along with a differentially suppressible tetracycline regulated expression system (DiSTRES). By differentially suppressing expression of the tetracycline-regulated transcriptional activator (tTA) and silencer (tTS) genes based on the cell origin, this leads to the activation and silencing of the TRE promoter, respectively. In vitro transduction of LNCaP cells with Ad/GFPDiSTRES lead to GFP expression levels that were over 30-fold higher than Ad/CMV-GFP. Furthermore, Ad/FasL-GFPDiSTRES demonstrated cytotoxic effects in prostate cancer cells known to be resistant to Fas-mediated apoptosis.
Conclusion:
Prostate-specific regulation from the DiSTRES system, therefore, serves as a promising new regulation strategy for future applications in the field of cancer gene therapy and gene therapy as a whole.
Insights
A new gene therapy strategy, DiSTRES, enhances cytotoxic gene expression in prostate cancer cells. This system overcomes weak promoter activity, offering a promising approach for improved cancer treatment outcomes.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Prostate cancer remains a leading cause of cancer death in American men, necessitating novel therapeutic strategies.
- Current gene therapy approaches for prostate cancer are limited by weak tissue-specific promoter activity, hindering effective transgene expression.
- Targeted delivery of cytotoxic genes offers a potential strategy to eliminate cancer cells while minimizing systemic toxicity.
Purpose of the Study:
- To develop an advanced gene expression regulation strategy for enhanced prostate-specific gene therapy.
- To overcome the limitations of weak promoter activity in tissue-specific gene expression for cancer treatment.
- To improve the efficacy of gene therapy for prostate cancer by amplifying cytotoxic gene delivery.
Main Methods:
- Development of a novel differentially suppressible tetracycline regulated expression system (DiSTRES).
- Incorporation of feedback amplification mechanisms to enhance gene expression levels.
- Utilizing cell-origin-based differential suppression of transcriptional activators (tTA) and silencers (tTS) to control TRE promoter activity.
Main Results:
- The DiSTRES system achieved over 30-fold higher GFP expression in LNCaP cells compared to standard CMV promoter systems.
- Demonstrated successful cytotoxic effects in prostate cancer cells resistant to Fas-mediated apoptosis using Ad/FasL-GFPDiSTRES.
- Successfully regulated gene expression in a prostate-specific manner via differential suppression of tTA and tTS.
Conclusions:
- The DiSTRES system represents a significant advancement in gene expression regulation for cancer gene therapy.
- This novel strategy shows promise for enhancing the efficacy of gene therapy in prostate cancer treatment.
- DiSTRES offers a versatile platform for future applications in gene therapy across various diseases.
