Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin

Shdema Filler-Hayut, Kiril Kniazev, Cathy Melamed-Bessudo*, Avraham A Levy*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Homologous recombination (HR) typically occurs during meiosis between homologs, at a few unplanned locations along the chromosomes. In this study, we tested whether targeted recombination between homologous chromosomes can be achieved via Clustered Regulatory Interspaced Short Palindromic Repeat associated protein Cas9 (CRISPR-Cas9)-induced DNA double-strand break (DSB) repair in Arabidopsis thaliana. Our experimental system includes targets for DSB induction in euchromatic and heterochromatic genomic regions of hybrid F1 plants, in one or both parental chromosomes, using phenotypic and molecular markers to measure Non-Homologous End Joining and HR repair. We present a series of evidence showing that targeted DSBs can be repaired via HR using a homologous chromosome as the template in various chromatin contexts including in pericentric regions. Targeted crossover was rare, but gene conversion events were the most frequent outcome of HR and were found in both “hot and cold” regions. The length of the conversion tracts was variable, ranging from 5 to 7505 bp. In addition, a typical feature of these tracks was that they often were interrupted. Our findings pave the way for the use of targeted gene-conversion for precise breeding.

Original languageEnglish
Article number12096
JournalInternational Journal of Molecular Sciences
Volume22
Issue number22
DOIs
Publication statusPublished - 9 Nov 2021

Funding

This research was funded by the Israel Science Foundation (ISF), grant 1027/14; and a Minerva grant to A.A.L and a donation from Erika Mogyoros.

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