Cleave and Rescue gamete killers create conditions for gene drive in plants
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1.
California Institute of Technology
Abstract
Gene drive elements promote the spread of linked traits and can be used to change the composition or fate of wild populations. Cleave and Rescue (ClvR) drive elements sit at a fixed chromosomal position and include a DNA sequence-modifying enzyme such as Cas9/gRNAs that disrupts endogenous versions of an essential gene and a recoded version of the essential gene resistant to cleavage. ClvR spreads by creating conditions in which those lacking ClvR die because they lack functional versions of the essential gene. Here we demonstrate the essential features of the ClvR gene drive in the plant Arabidopsis thaliana through killing of gametes that fail to inherit a ClvR that targets the essential gene YKT61. Resistant alleles, which can slow or prevent drive, were not observed. Modelling shows plant ClvRs are robust to certain failure modes and can be used to rapidly drive population modification or suppression. Possible applications are discussed.
Copyright and License
© The Author(s), under exclusive licence to Springer Nature Limited 2024.
Acknowledgement
We thank E. Meyerowitz and members of the Meyerowitz Lab Paul Tarr and Carla de Agostini Verna for introducing us to techniques for Arabidopsis maintenance, transgenesis and crossing. This work was supported by a grant to B.A.H. from the Caltech Center for Evolutionary Science (G.O. and M.L.J.) and the Caltech Resnick Sustainability Institute Explorer Grant (G.O.). T.I. was supported by NIH Training grant number 5T32GM007616-39 and with support to B.A.H. from the US Department of Agriculture, National Institute of Food and Agriculture (NIFA) specialty crop initiative under US Department of Agriculture NIFA award number 2012-51181-20086.
Contributions
Conceptualization, G.O., T.I. and B.A.H.; methodology, G.O., T.I., M.L.J., I.A. and B.A.H.; investigation, G.O., M.L.J., I.A. and B.A.H.; writing—original draft, G.O. and B.A.H.; writing—review and editing, G.O., T.I., M.L.J., I.A. and B.A.H.; funding acquisition, B.A.H.
Data Availability
All data are available in the main text and the Supplementary Information files. Illumina sequencing reads were deposited to SRA (bioproject, PRJNA1074841). The Arabidopsis TAIR 10 genome assembly was used in this study. Constructs and seeds of transgenic plants created in this study are available upon request.
Extended Data Fig. 2 T3 heterozygous ClvR crosses for (a) female ClvRap and (b) male ClvRap.
Code Availability
Modelling code and more information on the model, the scripts and parameters used to generate the data, and the data itself can be found at https://github.com/HayLab/Pigss. Plots were generated in R (version 4.2.3) with the ggplot2 package122.
Conflict of Interest
The authors have filed patent applications on ClvR and related technologies (US application numbers 15/970,728 and 16/673,823).
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Additional details
Identifiers
Related works
- Featured in
- Caltech News: https://www.caltech.edu/about/news/a-new-crispr-driven-technology-for-gene-drive-in-plants (URL)
- Is new version of
- Discussion Paper: 10.1101/2023.10.13.562303 (DOI)
- Discussion Paper: PMC10592828 (PMCID)
Funding
- California Institute of Technology
- Caltech Center for Evolutionary Science
- Resnick Sustainability Institute
- National Institutes of Health
- NIH Predoctoral Fellowship 5T32GM007616-39
- National Institute of Food and Agriculture
- 2012-51181-20086