Abstract
Plasmids currently used for nonviral gene transfer have the disadvantage of carrying a bacterial origin of replication and an antibiotic resistance gene. There is, therefore, a risk of uncontrolled dissemination of the therapeutic gene and the antibiotic resistance gene. Minicircles are new DNA delivery vehicles which do not have such elements and are consequently safer as they exhibit a high level of biological containment. They are obtained in E. coli by att site-specific recombination mediated by the phage λ integrase. The desired eukaryotic expression cassette bounded by the λ attP and attB sites was cloned on a recombinant plasmid. The expression cassette was excised in vivo after thermoinduction of the integrase gene leading to the formation of two supercoiled molecules: the minicircle and the starting plasmid lacking the expression cassette. In various cell lines, purified minicircles exhibited a two- to 10-fold higher luciferase reporter gene activity than the unrecombined plasmid. This could be due to either the removal of unnecessary plasmid sequences, which could affect gene expression, or the smaller size of minicircle which may confer better extracellular and intracellular bioavailability and result in improved gene delivery properties.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Darquet, AM., Cameron, B., Wils, P. et al. A new DNA vehicle for nonviral gene delivery: supercoiled minicircle. Gene Ther 4, 1341–1349 (1997). https://doi.org/10.1038/sj.gt.3300540
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300540
Keywords
This article is cited by
-
Considering the potential for gene-based therapy in prostate cancer
Nature Reviews Urology (2021)
-
CARAMBA: a first-in-human clinical trial with SLAMF7 CAR-T cells prepared by virus-free Sleeping Beauty gene transfer to treat multiple myeloma
Gene Therapy (2021)
-
Nanoparticles for drug delivery in Parkinson’s disease
Journal of Neurology (2021)
-
Highly branched poly(β-amino ester) delivery of minicircle DNA for transfection of neurodegenerative disease related cells
Nature Communications (2019)
-
A novel approach for assessment of prostate cancer aggressiveness using survivin-driven tumour-activatable minicircles
Gene Therapy (2019)