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Applications of and considerations for using CRISPR–Cas9-mediated gene conversion systems in rodents

Nature Protocols volume 17, pages 3–14 (2022)Cite this article

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Abstract

Genetic elements that are inherited at super-Mendelian frequencies could be used in a ‘gene drive’ to spread an allele to high prevalence in a population with the goal of eliminating invasive species or disease vectors. We recently demonstrated that the gene conversion mechanism underlying a CRISPR–Cas9-mediated gene drive is feasible in mice. Although substantial technical hurdles remain, overcoming these could lead to strategies that might decrease the spread of rodent-borne Lyme disease or eliminate invasive populations of mice and rats that devastate island ecology. Perhaps more immediately achievable at moderate gene conversion efficiency, applications in a laboratory setting could produce complex genotypes that reduce the time and cost in both dollars and animal lives compared with Mendelian inheritance strategies. Here, we discuss what we have learned from early efforts to achieve CRISPR–Cas9-mediated gene conversion, potential for broader applications in the laboratory, current limitations, and plans for optimizing this potentially powerful technology.

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Fig. 1: Self-propagating system versus split system.
Fig. 2: Meiotic timelines inform chromosome alignment.
Fig. 3: Active genetic strategies for the production of a variety of mouse models.
Fig. 4: Models of multilocus inheritance.

Data availability

No original data or code has been generated or used for this paper. Equations used for models are available in the text or figure legends.

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Acknowledgements

The authors acknowledge the input and advice of the Cooper laboratory as well as three referees who provided expert feedback and valuable clarifications for this work. This work was supported by National Institutes of Health R21GM129448 and a generous gift from the Tata Trusts in India to the Tata Institute for Genetics and Society–University of California San Diego (TIGS-UCSD) and TIGS-India. H.A.G. and A.J.W. were supported by National Institutes of Health training grant T32GM007240.

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  1. Division of Biological Sciences, Section of Cell and Developmental Biology, University of California San Diego, La Jolla, CA, USA

    Hannah A. Grunwald, Alexander J. Weitzel & Kimberly L. Cooper

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H.A.G., A.J.W. and K.L.C. conceptualized, wrote and edited the manuscript.

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Correspondence to Kimberly L. Cooper.

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K.L.C holds an advisory board position with Synbal, Inc. All other authors declare no competing interests.

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Grunwald, H. A. et al. Nature 566, 105–109 (2019): https://doi.org/10.1038/s41586-019-0875-2

Weitzel, A. J. et al. PLoS Biol. 19, e3001478 (2021): https://doi.org/10.1371/journal.pbio.3001478

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Grunwald, H.A., Weitzel, A.J. & Cooper, K.L. Applications of and considerations for using CRISPR–Cas9-mediated gene conversion systems in rodents. Nat Protoc 17, 3–14 (2022). https://doi.org/10.1038/s41596-021-00646-7

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