Megabulb DNA (mbDNA™) is a cell-free circular single-stranded DNA (ssDNA) template developed by Touchlight for non-viral gene editing and transient expression. Compared to linear ssDNA and double-stranded DNA (dsDNA), mbDNA offers higher homology-directed repair (HDR) efficiency, lower toxicity, and reduced immunogenicity. Its covalently closed design increases stability, enables nuclease complex formation, and enhances nuclear transport. In primary T cells, mbDNA achieves up to 70% CRISPR knock-in efficiency—matching viral vector performance—while supporting better cell recovery and expansion. Lipid nanoparticle delivery further mitigates electroporation toxicity. Capable of carrying multi-kilobase sequences without viral packaging limits, mbDNA is scalable to GMP-grade multi-milligram quantities. Leading industry evaluations in T cells and stem cells indicate broad potential for safer, efficient, and adaptable gene and cell therapy applications. More recently, mbDNA has shown to be a potent episomal expression vector, which promises reduced immunogenicity for in vivo non-viral gene therapy approaches. 

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