A temperature-sensitive and interferon-silent Sendai virus vector for CRISPR-Cas9 delivery and gene editing in primary human cells

Stevens, C. S.; Carmichael, J.; Watkinson, R.; Kowdle, S. S.; Reis, R. A.; Hamane, K.; Jang, J.; Park, A.; Pernet, O.; Khamaikawin, W.; Hong, P.; Thibault, P. A.; Gowlikar, A.; An, D. S.; Lee, B.

doi:10.1101/2024.05.03.592383

bioRxiv · 10.1101/2024.05.03.592383

A temperature-sensitive and interferon-silent Sendai virus vector for CRISPR-Cas9 delivery and gene editing in primary human cells

Stevens, C. S.·Carmichael, J.·Watkinson, R.·Kowdle, S. S.·Reis, R. A.·Hamane, K.·Jang, J.·Park, A.·Pernet, O.·Khamaikawin, W.·Hong, P.·Thibault, P. A.·Gowlikar, A.·An, D. S.·Lee, B.

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Abstract

The transformative potential of gene editing technologies hinges on the development of safe and effective delivery methods. In this study, we developed a temperature-sensitive and interferon-silent Sendai virus (ts SeV) as a novel delivery vector for CRISPR-Cas9 and for efficient gene editing in sensitive human cell types without inducing IFN responses. ts SeV demonstrates unprecedented transduction efficiency in human CD34+ hematopoietic stem and progenitor cells (HSPCs) including transduction of the CD34+/CD38-/CD45RA-/CD90+(Thy1+)/CD49fhigh stem cell enriched subpopulation. The frequency of CCR5 editing exceeded 90% and bi-allelic CCR5 editing exceeded 70% resulting in significant inhibition of HIV-1 infection in primary human CD14+ monocytes. These results demonstrate the potential of the ts SeV platform as a safe, efficient, and flexible addition to the current gene-editing tool delivery methods, which may help to further expand the possibilities in personalized medicine and the treatment of genetic disorders.

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Stevens, C. S., Carmichael, J., Watkinson, R., Kowdle, S. S., Reis, R. A., Hamane, K., Jang, J., Park, A., Pernet, O., Khamaikawin, W., Hong, P., Thibault, P. A., Gowlikar, A., An, D. S., Lee, B.. 2024-05-05. A temperature-sensitive and interferon-silent Sendai virus vector for CRISPR-Cas9 delivery and gene editing in primary human cells. https://doi.org/10.1101/2024.05.03.592383

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