Overriding Mendelian inheritance inArabidopsiswith a CRISPR toxin-antidote gene drive that impairs pollen germination

Abstract

AbstractSynthetic gene drives, inspired by natural selfish genetic elements, present transformative potential for disseminating traits that benefit humans throughout wild populations, irrespective of potential fitness costs. Here, we constructed a gene drive system called CRISPR-Assisted Inheritance utilizingNPG1(CAIN), which employs a toxin-antidote mechanism in the male germline to override Mendelian inheritance in plants. Specifically, a gRNA-Cas9 cassette targets the essentialNo Pollen Germination 1(NPG1) gene, serving as the toxin to block pollen germination. A recoded, CRISPR-resistant copy ofNPG1serves as the antidote, providing rescue only in pollen cells that carry the drive. To limit potential consequences of inadvertent release, we used self-pollinatingArabidopsis thalianaas a model. The drive demonstrated a robust 88–99% transmission rate over two successive generations, producing minimal resistance alleles that are unlikely to inhibit drive spread. Our study provides a strong basis for rapid genetic modification or suppression of outcrossing plant populations.