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Updated: Jan 9, 2026

Production of Double-stranded DNA Ministrings

Production of Double-stranded DNA Ministrings

Published on: February 29, 2016

Plasmid DNA Delivery Using a Stable Nanovesicle Platform: A Design-of-Experiments-Guided Investigation.

Mariana Köber^1,2, Irene González-Domínguez³, Diego Valdospinos^1,3

¹Institute of Materials Science of Barcelona (ICMAB-CSIC), Universitat Autònoma de Barcelona, Cerdanyola del Vallès, 08193 Barcelona, Spain.

ACS Biomaterials Science & Engineering

|December 5, 2025

Summary

This summary is machine-generated.

Stable lipid-based nanovesicles effectively deliver plasmid DNA (pDNA) into cells. Optimization of parameters like DNA concentration and lipid ratios is crucial for efficient nonviral gene delivery, advancing biotechnological applications.

Keywords:

Design of Experiments HEK293 cell transfection green fluorescent protein lipid-based nanovesicles nonviral vectors plasmid DNA delivery quatsomes

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Area of Science:

Biotechnology
Nanomedicine
Molecular Biology

Background:

Plasmid DNA (pDNA) delivery is vital for biotechnology and medicine.
Nonviral lipid-based nanocarriers show promise for mammalian cell transfection.
Developing broadly effective and scalable DNA delivery systems remains a challenge.

Purpose of the Study:

To evaluate stable nanovesicles of cholesteryl N-(2-dimethylaminoethyl)carbamate (DC-CHOL) and myristalkonium chloride (MKC) for pDNA delivery.
To optimize transfection parameters using a Design of Experiments (DoE) approach.
To assess the suitability of these nanovesicles for large-scale biotherapeutic production.

Main Methods:

Utilized DC-CHOL/MKC nanovesicles for plasmid DNA delivery.
Employed suspension-adapted human embryonic kidney 293 (HEK293) cells.
Applied a Design of Experiments (DoE) approach to optimize DNA concentration, DNA-to-vesicle ratio, and NaCl concentration.

Main Results:

DC-CHOL/MKC nanovesicles demonstrated efficient delivery of double-stranded plasmid DNA.
Optimized conditions significantly improved transfection efficiency.
Physicochemical properties of the pDNA/lipid complex were found to be critical for transfection.

Conclusions:

DC-CHOL/MKC nanovesicles are effective nonviral vectors for pDNA delivery.
Parameter optimization via DoE is essential for maximizing transfection efficiency.
These nanovesicles hold potential for scalable gene delivery in biopharmaceutical applications.