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Review
. 2024 Mar 12;121(11):e2307796121.
doi: 10.1073/pnas.2307796121. Epub 2024 Mar 4.

Targeted nonviral delivery of genome editors in vivo

Connor A Tsuchida #  1   2 , Kevin M Wasko #  2   3 , Jennifer R Hamilton #  2   3 , Jennifer A Doudna  1   2   3   4   5   6   7   8
Affiliations
Review

Targeted nonviral delivery of genome editors in vivo

Connor A Tsuchida et al. Proc Natl Acad Sci U S A. .

Abstract

Cell-type-specific in vivo delivery of genome editing molecules is the next breakthrough that will drive biological discovery and transform the field of cell and gene therapy. Here, we discuss recent advances in the delivery of CRISPR-Cas genome editors either as preassembled ribonucleoproteins or encoded in mRNA. Both strategies avoid pitfalls of viral vector-mediated delivery and offer advantages including transient editor lifetime and potentially streamlined manufacturing capability that are already proving valuable for clinical use. We review current applications and future opportunities of these emerging delivery approaches that could make genome editing more efficacious and accessible in the future.

Keywords: CRISPR-Cas; genome editing; in vivo delivery; nonviral delivery; targeted delivery.

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Conflict of interest statement

Competing interests statement:J.A.D. is a co-founder of Editas Medicine, Intellia Therapeutics, Caribou Biosciences, Mammoth Biosciences, and Scribe Therapeutics. J.R.H. and J.A.D. are co-founders of Azalea Therapeutics. J.A.D is a scientific advisory board member of Intellia Therapeutics, Caribou Biosciences, Mammoth Biosciences, Scribe Therapeutics, Vertex Pharmaceuticals, eFFECTOR Therapeutics, Felix Biosciences, The Column Group, Inari, and Isomorphic Labs. J.A.D. is the Chief Science Advisor at Sixth Street, an advisor at Tempus, and a Director at Johnson & Johnson and Altos Labs. The Regents of the University of California have patents issued and/or pending related to CRISPR technologies and delivery technologies on which C.A.T., J.R.H., and J.A.D. are listed inventors. J.A.D. has sponsored research projects through Biogen, Pfizer, Apple Tree Partners, Genentech, and Roche.

Figures

Fig. 1.
Fig. 1.
Preclinical and clinical delivery methods for nonviral CRISPR-Cas genome editing. The delivery of CRISPR-Cas RNP and mRNA has been a major focus of the field. Techniques such as ex vivo delivery to T cells and HSPCs, ex vivo delivery to iPSC-derived pancreatic cells, and systemic delivery to the liver have already reached the clinic (highlighted with asterisks). While continued research efforts have developed limited delivery methods for tissues such as the lung and spleen, systemic delivery methods to the brain, heart, HSPCs, and muscle (highlighted with exclamation marks) remain a major unmet need and are poised to become the focus of future work. This image utilized assets from freepik.com.
See this image and copyright information in PMC

References

    1. Gaj T., Gersbach C. A., Barbas C. F., ZFN, TALEN and CRISPR/Cas-based methods for genome engineering. Trends Biotechnol. 31, 397–405 (2013). - PMC - PubMed
    1. Adli M., The CRISPR tool kit for genome editing and beyond. Nat. Commun. 9, 1911 (2018). - PMC - PubMed
    1. Tsai S. Q., et al. , GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases. Nat. Biotechnol. 33, 187–197 (2015). - PMC - PubMed
    1. Haeussler M., et al. , Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR. Genome Biol. 17, 148 (2016). - PMC - PubMed
    1. Kim N., et al. , Prediction of the sequence-specific cleavage activity of Cas9 variants. Nat. Biotechnol. 38, 1328–1336 (2020). - PubMed