ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Plant Cell Biology
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1580225
This article is part of the Research TopicMeiotic Recombination and DNA Repair: New Approaches to Solve Old Questions in Model and Non-Model Plant Species - Volume IIView all articles
dCas9-SPO11-1 locally stimulates meiotic recombination in rice
Provisionally accepted
Léo Herbert1
Aurore Vernet2
Julien Frouin2
Anne Cécile Meunier2Jeremy Di Mattia3Minghui Wang4Gaganpreet K. Sidhu4Luc Mathis4Alain Nicolas4*Emmanuel Guiderdoni2*Ian Fayos1*- 1Meiogenix, Paris, France
- 2CIRAD UMR AGAP, Montpellier, France
- 3IAGE FRANCE, Montpellier, France
- 4Meiogenix Inc., Ithaca, New York, United States
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Meiotic crossovers shuffle the genetic information transmitted by the gametes. However, the potential to recover all the combinations of the parental alleles remains limited in most organisms, including plants, by the occurrence of only few crossovers per chromosome and a prominent bias in their spatial distribution. Thus, novel methods for stimulating recombination frequencies and/or modifying their location are highly desired to accelerate plant breeding. Here, we investigate the use of a dCas9-SPO11-1 fusion and clusters of 11 gRNAs to alter meiotic recombination in two chromosomal regions of a rice hybrid (KalingaIII/Kitaake). Expression of the dCas9-SPO11-1 fusion protein under the ubiquitous ZmUbi1 promoter was obtained in leaves/anthers/meiocytes and found to complement the sterility of the Osspo11-1 mutant line. To accurately genotype rare recombinants in regions of few kbp, we improved the digital PCR-based pollen-typing method and observed a 3.27-fold increase over wild-type (p<0.001) of recombinant pollens in a transgenic hybrid line (7a) targeting a chromosome 7 region. In the offspring plant 7a1, a significant 2.05-fold increase (p=0.048) was observed in the central interval (7.2 kb) of the Chr. 7 target region. This stimulation of meiotic recombination is consistent with the expression of the dCas9-SPO11-1 fusion and gRNAs as well as with the ChIP-revealed binding of dCas9-SPO11-1 to the targeted region. In contrast, no stimulation was observed in other transgenic lines deficient in the above pre-requisite features, expressing the dCas9-SPO11-1 fusion but no gRNAs or targeting a Chr.9 region. These results open new avenues to locally stimulate meiotic recombination in crop genomes and paves the way for a future implementation in plant breeding programs.
Keywords: CRISPR/dCas9, Meiosis, plant breeding, pollen typing, rice, Spo11, Targeted recombination
Received: 20 Feb 2025; Accepted: 31 Mar 2025.
Copyright: © 2025 Herbert, Vernet, Frouin, Meunier, Di Mattia, Wang, Sidhu, Mathis, Nicolas, Guiderdoni and Fayos. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Alain Nicolas, Meiogenix Inc., Ithaca, 14853, New York, United States
Emmanuel Guiderdoni, CIRAD UMR AGAP, Montpellier, France
Ian Fayos, Meiogenix, Paris, 75011, France
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