From Plasmids To Cell-Free DNA For Advanced Gene Therapies

By Elena Stoyanova, PhD. Principle Scientist, Platform Research, Touchlight and Chesney Michels, PhD. Director, Emerging Technology for Cell and Gene Therapy, Elevate Bio

The evolution of genetic medicines is driving a significant shift from viral vectors toward non-viral gene modification technologies to address safety and scalability concerns. Traditional methods relying on bacterial plasmid DNA fermentation introduce unwanted sequence elements and often result in high cellular toxicity. Transitioning to synthetic, cell-free circular single-stranded DNA platforms provides a streamlined alternative specifically optimized as templates for homology-directed repair.

Eliminating bacterial-derived components minimizes innate immunogenicity, substantially improving cell viability and recovery across sensitive primary immune and stem cell populations. Real-world applications demonstrate that these minimal, user-defined circular architectures can achieve up to 75% knock-in efficiency while optimizing downstream cellular yields. Embracing advanced enzymatic architectures allows developers to bypass long lead times and scale non-viral therapeutic pipelines with greater precision.