An Autogenously Regulated Expression System for Gene Therapeutic Ocular Applications

Area of Science:

• Molecular Biology
• Gene Therapy
• Biotechnology

Background:

• Effective gene therapy requires precise control over transgene expression.
• Existing methods for regulating transgenes in vivo can be limited.

Purpose of the Study:

• To develop and validate a novel, inducible transgene regulatory system for controlling gene expression in various cell types and in vivo.
• To assess the efficacy and reversibility of the autogenous transgene regulatory system (ARES) in the mouse retina.

Main Methods:

• Development of the autogenous transgene regulatory system (ARES) based on the bacterial lac repressor.
• Testing ARES in bacteria and eukaryotic cells.
• In vivo studies using adeno-associated virus (AAV) vectors in mouse retina, with transgene expression regulated by Isopropyl β-D-1-thiogalactopyranoside (IPTG).

Main Results:

• The ARES system demonstrated successful control of transgene expression in bacteria, eukaryotic cells, and mouse retina.
• Luciferase expression in the mouse retina was reversibly controlled over three induction-repression cycles via oral IPTG administration.
• IPTG, a non-pharmacologic molecule, showed no off-target effects in mammals.

Conclusions:

• The autogenous transgene regulatory system (ARES) offers a robust and reversible method for controlling transgene expression.
• Oral administration of IPTG for in vivo transgene induction presents a promising strategy for future gene therapy applications.
• This system holds significant potential for treating inherited and acquired genetic diseases.