Advanced Peptide Nanoparticles Enable Robust and Efficient delivery of gene editors across cell types

Gustafsson, O.; Krishna, S.; Borate, S.; Ghaeidamini, M.; Liang, X.; Saher, O.; Cuellar, R.; Birdsong, B. K.; Roudi, S.; Estupinan, Y. H.; Alici, E.; Smith, E. C.; Esbjörner, E. K.; Spuler, S. K.; de Jong, O. G.; Escobar, H.; Nordin, J. Z.; Andaloussi, S. E.

doi:10.1101/2024.11.27.624305

bioRxiv · 10.1101/2024.11.27.624305

Advanced Peptide Nanoparticles Enable Robust and Efficient delivery of gene editors across cell types

Gustafsson, O.·Krishna, S.·Borate, S.·Ghaeidamini, M.·Liang, X.·Saher, O.·Cuellar, R.·Birdsong, B. K.·Roudi, S.·Estupinan, Y. H.·Alici, E.·Smith, E. C.·Esbjörner, E. K.·Spuler, S. K.·de Jong, O. G.·Escobar, H.·Nordin, J. Z.·Andaloussi, S. E.

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Abstract

Efficient delivery of the CRISPR/Cas9 system and its larger derivatives, base editors, and prime editors remain a significant challenge, particularly in tissue-specific stem cells and induced pluripotent stem cells (iPSCs). This study optimized a novel family of cell-penetrating peptides, hPep, to deliver gene-editing ribonucleoproteins. The hPep-based nanoparticles enable highly efficient and biocompatible delivery of Cre recombinase, Cas9, base-, and prime editors. Using base editors, robust and nearly complete genome editing was achieved in the human cells: HEK293T (96%), iPSCs (74%), and muscle stem cells (80%). This strategy opens promising avenues for ex vivo and, potentially, in vivo applications. Incorporating silica particles enhanced the systems versatility, facilitating cargo-agnostic delivery. Notably, the nanoparticles can be synthesized quickly on a benchtop and stored as lyophilized powder without compromising functionality. This represents a significant advancement in the feasibility and scalability of gene-editing delivery technologies.

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Gustafsson, O., Krishna, S., Borate, S., Ghaeidamini, M., Liang, X., Saher, O., Cuellar, R., Birdsong, B. K., Roudi, S., Estupinan, Y. H., Alici, E., Smith, E. C., Esbjörner, E. K., Spuler, S. K., de Jong, O. G., Escobar, H., Nordin, J. Z., Andaloussi, S. E.. 2024-12-02. Advanced Peptide Nanoparticles Enable Robust and Efficient delivery of gene editors across cell types. https://doi.org/10.1101/2024.11.27.624305

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