Plant Gene Drive Tool - CAIN

On June 17, 2024, Qian Wenfeng's team from the Chinese Academy of Sciences published a research paper titled "Overriding Mendelian inheritance in Arabidopsis with a CRISPR toxin-antidote gene drive that impairs pollen germination" in Nature Plants.

This study reports an artificial gene drive system, CRISPR-Assisted Inheritance utilizing NPG1 (CAIN), which can lead to the generational advantage of CAIN-carrying pollen, thereby realizing plant super-Mendelian inheritance and laying the foundation for genetic manipulation and population suppression of natural plant populations. To ensure ecological safety, the researchers conducted a proof-of-concept design of the system in self-propagated Arabidopsis thaliana.

Faced with challenges such as the threat posed by weeds to agricultural production and the environmental crisis caused by invasive plants, genetic control of wild plants at the population level has become a promising strategy. However, there is a type of selfish genes or genetic elements in plant genomes, which are passed on to future generations at a rate that exceeds Mendel's laws. They are called gene drive elements. Inspired by natural gene drive elements, the development of artificial gene drive tools provides a potential solution for modifying wild plant populations.

Qian Wenfeng's team used CRISPR technology to develop the plant artificial gene drive system CAIN. This system is based on the "toxin-antidote" mechanism, using the CRISPR/Cas9 component that cuts the gene No Pollen Germination 1 (NPG1) necessary for Arabidopsis pollen germination as a "toxin" to prevent pollen germination. At the same time, the NPG1 sequence recoded using synonymous codons and not cleaved by CRISPR/Cas9 is used as an antidote, thereby replenishing the gene functions required for normal germination of pollen carrying gene drive elements.

When a heterozygous plant carrying CAIN is used as the male parent, because two copies of endogenous NPG1 are knocked out, only pollen carrying CAIN can germinate successfully, and the theoretical transmission rate reaches 100%. It was verified by artificial hybridization experiments that the male parent plant carrying CAIN showed significant supra-Mendelian genetic effects in two consecutive generations of hybridization with wild-type plants. The CAIN transmission rate is 88-99%, which is much higher than the 50% expected by Mendelian inheritance.

Because NPG1 is conserved among species, or other related genes that control pollen germination can be used as alternative target genes, this system provides a way to achieve population modification or population suppression of natural plant populations. Through design optimization, plant gene drive tools are expected to play an important role in future agricultural weed control and ecological protection.

For example, CAIN is inserted into a herbicide resistance gene to destroy the function of the gene. Through super-Mendelian inheritance of CAIN, farmland weed populations can recover their sensitivity to herbicides after multiple generations of hybridization. In addition, CAIN can be used to control the population size of invasive plants by destroying fertility-related genes. Or, endangered plants can be saved by introducing stress-resistant genes.

However, gene drive tools are a double-edged sword. How to control the intensity, spatial and temporal scope, and species specificity of genetic manipulation of outcrossed species in the wild, as well as to prevent malicious releases, urgently requires the participation, discussion, and formulation of management regulations by the scientific community, policymakers, and stakeholders.

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Reference

1. Liu, Y., et al. Overriding Mendelian inheritance in Arabidopsis with a CRISPR toxin–antidote gene drive that impairs pollen germination. Plants. 10, 910–922 (2024).