dCas9-SPO11-1 locally stimulates meiotic recombination in rice
IntroductionMeiotic 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...
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Frontiers Media S.A.
2025-05-01
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| Series: | Frontiers in Plant Science |
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| author | Léo Herbert Aurore Vernet Aurore Vernet Julien Frouin Julien Frouin Anne Cécile Meunier Anne Cécile Meunier Jeremy Di Mattia Minghui Wang Gaganpreet K. Sidhu Luc Mathis Alain Nicolas Alain Nicolas Emmanuel Guiderdoni Emmanuel Guiderdoni Ian Fayos |
| author_facet | Léo Herbert Aurore Vernet Aurore Vernet Julien Frouin Julien Frouin Anne Cécile Meunier Anne Cécile Meunier Jeremy Di Mattia Minghui Wang Gaganpreet K. Sidhu Luc Mathis Alain Nicolas Alain Nicolas Emmanuel Guiderdoni Emmanuel Guiderdoni Ian Fayos |
| author_sort | Léo Herbert |
| collection | DOAJ |
| description | IntroductionMeiotic 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. MethodsHere, 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). To accurately genotype rare recombinants in regions of few kbp, we improved the digital PCR-based pollen-typing method in parallel.ResultsExpression 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. We 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. DiscussionThese results open new avenues to locally stimulate meiotic recombination in crop genomes and paves the way for a future implementation in plant breeding programs. |
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| publishDate | 2025-05-01 |
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| spelling | doaj-art-1b50f8f7cce54e97a6863b07fdc22e0c2025-08-20T01:48:26ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-05-011610.3389/fpls.2025.15802251580225dCas9-SPO11-1 locally stimulates meiotic recombination in riceLéo Herbert0Aurore Vernet1Aurore Vernet2Julien Frouin3Julien Frouin4Anne Cécile Meunier5Anne Cécile Meunier6Jeremy Di Mattia7Minghui Wang8Gaganpreet K. Sidhu9Luc Mathis10Alain Nicolas11Alain Nicolas12Emmanuel Guiderdoni13Emmanuel Guiderdoni14Ian Fayos15Meiogenix SA, Paris, FranceCentre de coopération internationale en recherche agronomique pour le développement (CIRAD), Unité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Montpellier, FranceUnité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Université de Montpellier, Centre de coopération internationale en recherche agronomique pour le développement (CIRAD), Institut national de recherche pour l'agriculture, l'alimentation et l'environnement (INRAE), Institut Agro, Montpellier, FranceCentre de coopération internationale en recherche agronomique pour le développement (CIRAD), Unité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Montpellier, FranceUnité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Université de Montpellier, Centre de coopération internationale en recherche agronomique pour le développement (CIRAD), Institut national de recherche pour l'agriculture, l'alimentation et l'environnement (INRAE), Institut Agro, Montpellier, FranceCentre de coopération internationale en recherche agronomique pour le développement (CIRAD), Unité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Montpellier, FranceUnité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Université de Montpellier, Centre de coopération internationale en recherche agronomique pour le développement (CIRAD), Institut national de recherche pour l'agriculture, l'alimentation et l'environnement (INRAE), Institut Agro, Montpellier, FranceIngénierie et Analyse en Génétique Environnementale (IAGE), Montpellier, FranceMeiogenix Inc., Center for Life Science Ventures Cornell University, Ithaca, NY, United StatesMeiogenix Inc., Center for Life Science Ventures Cornell University, Ithaca, NY, United StatesMeiogenix SA, Paris, FranceMeiogenix SA, Paris, FranceIRCAN (Institute for Research on Cancer and Aging), CNRS (Centre national de la recherche scientifique) UMR7284, INSERM (Institut national de la santé et de la recherche médicale) U1081, Université Côte d’Azur, Nice, FranceCentre de coopération internationale en recherche agronomique pour le développement (CIRAD), Unité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Montpellier, FranceUnité mixte de recherche - Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) Institut, Université de Montpellier, Centre de coopération internationale en recherche agronomique pour le développement (CIRAD), Institut national de recherche pour l'agriculture, l'alimentation et l'environnement (INRAE), Institut Agro, Montpellier, FranceMeiogenix SA, Paris, FranceIntroductionMeiotic 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. MethodsHere, 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). To accurately genotype rare recombinants in regions of few kbp, we improved the digital PCR-based pollen-typing method in parallel.ResultsExpression 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. We 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. DiscussionThese results open new avenues to locally stimulate meiotic recombination in crop genomes and paves the way for a future implementation in plant breeding programs.https://www.frontiersin.org/articles/10.3389/fpls.2025.1580225/fullCRISPR/dCas9meiosisplant breedingpollen typingriceSpo11 |
| spellingShingle | Léo Herbert Aurore Vernet Aurore Vernet Julien Frouin Julien Frouin Anne Cécile Meunier Anne Cécile Meunier Jeremy Di Mattia Minghui Wang Gaganpreet K. Sidhu Luc Mathis Alain Nicolas Alain Nicolas Emmanuel Guiderdoni Emmanuel Guiderdoni Ian Fayos dCas9-SPO11-1 locally stimulates meiotic recombination in rice Frontiers in Plant Science CRISPR/dCas9 meiosis plant breeding pollen typing rice Spo11 |
| title | dCas9-SPO11-1 locally stimulates meiotic recombination in rice |
| title_full | dCas9-SPO11-1 locally stimulates meiotic recombination in rice |
| title_fullStr | dCas9-SPO11-1 locally stimulates meiotic recombination in rice |
| title_full_unstemmed | dCas9-SPO11-1 locally stimulates meiotic recombination in rice |
| title_short | dCas9-SPO11-1 locally stimulates meiotic recombination in rice |
| title_sort | dcas9 spo11 1 locally stimulates meiotic recombination in rice |
| topic | CRISPR/dCas9 meiosis plant breeding pollen typing rice Spo11 |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2025.1580225/full |
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