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  • Perspective
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Prototype mouse models for researching SEND-based mRNA delivery and gene therapy

Nature Protocols volume 17, pages 2129–2138 (2022)Cite this article

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Abstract

One of the major challenges of gene therapy—an approach to treat diseases caused by faulty genes—is a lack of technologies that deliver healthy gene copies to target tissues and cells. Some commonly used approaches include viral vectors or coating therapeutic nucleic acids with lipid-based nanoparticles to pass through cell membranes, but these technologies have had limited success. A revolutionary tool, the CRISPR–Cas gene-editing system, offers tremendous promise, but it too suffers from problems with delivery. Another tool, called ‘SEND’ (for ‘selective endogenous encapsidation for cellular delivery’), seems to offer a better solution. The SEND system uses endogenous genetic components to package mRNA cargoes to deliver them to other cells via virus-like particles (VLPs). The SEND-VLP tool has enormous potential as a gene-therapy tool, if the endogenous components of SEND can be repurposed to produce VLPs containing therapeutic cargoes. However, several aspects of this newly identified phenomenon are not yet fully understood. Genetically engineered mouse (GEM) models, expressing different combinations of SEND components in a controllable and inducible fashion, could serve as valuable tools to understand more about this tool and to repurpose it for gene-therapy applications. In this Perspective, we discuss how GEM models and mouse molecular genetics tools could be used for SEND-VLP research.

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Fig. 1: Schematic of SEND-VLP concept.
Fig. 2: Non-GEM strategies for testing the SEND system in mice.
Fig. 3: GEM strategies for testing the SEND system in mice.

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No original data or code was generated or used for this paper.

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Acknowledgements

We thank Guy Richardson (University of Sussex), Suzanne Mansour (University of Utah) and Hiromi Miura (Tokai University) for critical reading of the manuscript and Nick May (TypeRight) and James M. Burbach (University of Nebraska Medical Center) for copy editing. C.B.G. is funded by NIH grants R35HG010719, R21GM129559, R21AI143394 and R21DA046831. M.O. is funded by JSPS KAKENHI (20K21551) and Takeda Science Foundation (2020).

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Authors and Affiliations

  1. Mouse Genome Engineering Core Facility, University of Nebraska Medical Center, Omaha, NE, USA

    Channabasavaiah B. Gurumurthy & Rolen M. Quadros

  2. Genome Editing and Education Center Nebraska (GEEC-Nebraska), College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA

    Channabasavaiah B. Gurumurthy & Rolen M. Quadros

  3. Department of Pharmacology and Experimental Neuroscience, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA

    Channabasavaiah B. Gurumurthy & Rolen M. Quadros

  4. Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Japan

    Masato Ohtsuka

  5. The Institute of Medical Sciences, Tokai University, Isehara, Japan

    Masato Ohtsuka

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  1. Channabasavaiah B. Gurumurthy
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Contributions

C.B.G. and M.O. conceived the overall idea and wrote the manuscript. R.M.Q. generated a figure schematic.

Corresponding authors

Correspondence to Channabasavaiah B. Gurumurthy or Masato Ohtsuka.

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Competing interests

A provisional patent on some of the ideas in this article will be submitted in which C.B.G. and M.O. will be listed as inventors.

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Nature Protocols thanks Rosie Bunton-Stasyshyn, Xucheng Hou and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Additional initial assessment was performed by informal referee Horacio Cabral.

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Key references using this protocol

Segel, M. et al. Science 373, 882–889 (2021); https://doi.org/10.1126/science.abg6155

Quadros, R. M. et al. Genome Biol. 18, 92 (2017); https://doi.org/10.1186/s13059-017-1220-4

Miura, H. et al. Nat. Protoc. 13, 195–215 (2018); https://doi.org/10.1038/nprot.2017.153

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Gurumurthy, C.B., Quadros, R.M. & Ohtsuka, M. Prototype mouse models for researching SEND-based mRNA delivery and gene therapy. Nat Protoc 17, 2129–2138 (2022). https://doi.org/10.1038/s41596-022-00721-7

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