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Constitutive and Inducible Systems for Genetic In Vivo Modification of Mouse Hepatocytes Using Hydrodynamic Tail Vein Injection
Published on: February 2, 2018
A designed curved DNA sequence remarkably enhances transgene expression from plasmid DNA in mouse liver
S Fukunaga1, G Kanda, J Tanase
1Hokkaido University, Sapporo, Japan.
Gene Therapy
|September 16, 2011
Summary
Adding a specific curved DNA sequence to plasmids significantly boosts transgene expression in vivo. This sequence facilitates histone interactions, promoting gene activity and offering a new method for controlling gene delivery.
Area of Science:
- Molecular Biology
- Gene Therapy
- Biochemistry
Background:
- Intranuclear plasmid DNA disposition is critical for effective transgene expression.
- Plasmid DNA and histone protein interactions are key regulators of DNA disposition and gene expression.
Purpose of the Study:
- To investigate the impact of a left-handedly curved DNA sequence on transgene expression in vivo.
- To explore the mechanism by which curved DNA sequences influence nucleosome positioning and transcriptional activity.
Main Methods:
- Constructed a naked luciferase plasmid incorporating a left-handedly curved sequence (20-40 repeated A•T tracts).
- Delivered the plasmid into mouse liver using hydrodynamics-based injection.
- Quantitated luciferase activity over time and analyzed nucleosome positioning.
Main Results:
- The inclusion of the curved DNA sequence markedly increased transgene expression.
- Analysis indicated the curved sequence acts as a histone core acceptor, facilitating nucleosome sliding.
- This nucleosome sliding was linked to transcriptional activation.
Conclusions:
- Designed curved DNA sequences can effectively control transgene expression from plasmid DNA in vivo.
- The findings suggest a novel strategy for enhancing gene delivery and expression efficiency using DNA structural elements.
