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Minicircle DNA (mcDNA), as a novel non-viral gene vector, has become an indispensable tool in gene therapy, cell therapy, and vaccine development, distinguished by its efficiency, safety, and long-term expression characteristics. Creative Biogene leverages a comprehensive technical platform with extensive industry experience to provide customized mcDNA products and services, driving advancements in gene therapy, cell therapy, and vaccine development. Whether for small-scale research or large-scale production, we deliver high-quality mcDNA products and technical support to accelerate research and clinical translation.
Minicircle DNA was first described in 1997 as a non-viral, extrachromosomal, covalently closed circular gene expression vector. It can carry target genes with minimal base pairs, making them the smallest DNA vector unit required for gene expression in microorganisms. It features a minimalist backbone that meets clinical therapy safety requirements and enables sustained gene expression, demonstrating enhanced transgene expression levels and safety in cellular environments. Furthermore, mcDNA typically does not integrate into the genome and maintains high safety in both quiescent and actively dividing cells, making it an ideal non-viral vector.
Figure 1. Schematic representation of in vivo parental plasmid intramolecular recombination.
McDNA is generated through the recombination of specialized parental plasmids in competent bacteria under induction. The parental plasmid contains attB and attP recombination sites and multiple restriction sites. When specific recombinase and endonuclease are produced under inducer action, the parental plasmid recombines to generate the minicircle plasmid (containing only the target sequence) and bacterial backbone plasmid (miniplasmid). The bacterial backbone plasmid is further degraded by endonucleases, yielding pure mcDNA.
Technical Advantages
Creative Biogene specializes in providing comprehensive mcDNA synthesis and manufacturing services. We have established GMP-compliant platforms to support the production of mcDNA. With extensive experience in process development and GMP manufacturing, we offer integrated services including mcDNA vector design, synthesis, optimization, quality control, stability testing, and GMP production of both non-clinical and clinical-grade mcDNA.
Customization Process
1. Gene Cloning
The target gene is cloned into a suitable parental plasmid containing essential sequences and promoters (e.g., CMV or EF1). Optimal gene sequences are selected to ensure efficient expression and functionality.
2. McDNA Production
Inducible E. coli strains are used for minicircle production, employing advanced recombinase systems (e.g., λ-integrase, Cre-recombinase, or ϕC31-integrase) to minimize multimeric forms and maximize pure minicircle DNA yield.
3. Purification
After bacterial lysis, minicircles are separated from residual miniplasmids and un-recombined parental plasmids through specialized purification methods, including hydrophobic interaction, anion exchange, and multimodal chromatography. This ensures high-purity mcDNA, free from contaminants, suitable for transfection and functional testing.
4. Quality Control
DNA electrophoresis and restriction enzyme analysis are performed to verify the purity, integrity, and identity of the minicircle DNA, ensuring compliance with downstream application requirements.
5. Gene Transfection and Application (If required)
For transfection experiments, the purified mcDNA undergoes further processing to eliminate any residual parental plasmid or genetic background contamination, ensuring high purity and functionality for its intended application. Rigorous quality control is maintained throughout the process.
Service Features
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