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Coexpression of MEIOTIC-TOPOISOMERASE VIB-dCas9 with guide RNAs specific to a recombination hotspot is insufficient to increase crossover frequency in Arabidopsis.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Holland, Daniel 
Gonzalez-Jorge, Sabrina  ORCID logo  https://orcid.org/0000-0002-0783-1929

Abstract

During meiosis, homologous chromosomes pair and recombine, which can result in reciprocal crossovers that increase genetic diversity. Crossovers are unevenly distributed along eukaryote chromosomes and show repression in heterochromatin and the centromeres. Within the chromosome arms, crossovers are often concentrated in hotspots, which are typically in the kilobase range. The uneven distribution of crossovers along chromosomes, together with their low number per meiosis, creates a limitation during crop breeding, where recombination can be beneficial. Therefore, targeting crossovers to specific genome locations has the potential to accelerate crop improvement. In plants, meiotic crossovers are initiated by DNA double-strand breaks that are catalyzed by SPO11 complexes, which consist of 2 catalytic (SPO11-1 and SPO11-2) and 2 noncatalytic subunits (MTOPVIB). We used the model plant Arabidopsis thaliana to coexpress an MTOPVIB-dCas9 fusion protein with guide RNAs specific to the 3a crossover hotspot. We observed that this was insufficient to significantly change meiotic crossover frequency or pattern within 3a. We discuss the implications of our findings for targeting meiotic recombination within plant genomes.

Description

Keywords

CRISPR/Cas9, MTOPVIB, crossover, meiosis, targeted recombination, Arabidopsis, Arabidopsis Proteins, Crossing Over, Genetic, Homologous Recombination, Meiosis, Plant Breeding, Plants, RNA, Guide, CRISPR-Cas Systems

Journal Title

G3 (Bethesda)

Conference Name

Journal ISSN

2160-1836
2160-1836

Volume Title

Publisher

Oxford University Press (OUP)
Sponsorship
Gatsby Charitable Foundation (GAT3587)
Biotechnology and Biological Sciences Research Council (BB/N007557/1)
BBSRC, Broodbank Trust, Gatsby Charitable Foundation