Background: Stable gene insertion using non-viral delivery is
essential to unlock the next generation of cell and gene therapies
that are both safer and and potentially more scalable, helping
reduce barriers to patient access. Currently, viral vectors carry
high manufacturing costs and insertional mutagenesis cocerns,
whereas electroporation (EP) affects cell yield and viability.
Non-viral lipid nanoparticles (LNPs) offer an alternative for gene
modification of T cells, enabling delivery that preserves cell
health, uses chemically defined manufacturing, and supports
scalable production.
Methods: Primary human T cells were edited using Cytiva’s T cell
Kit through the co-delivery of Cas9 mRNA, sgRNA, and a
single-stranded oligodeoxynucleotide (ssODN) donor (HA tag at
CD5). Outcomes were assessed by flow cytometry 4 days
post-treatment.
Results: Across 7 donors, LNPs achieved LNPs achieve 31 ± 7%
homology-directed repair (HDR) (day 4) with 96 ± 5% relative
viability, n=7 donors.
Conclusions: LNPs enable high efficiency and consistent ssODN
HDR in primary T cells supporting non-viral approaches for cell
therapy research.