Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin

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 ass...

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Autores principales:	Shdema Filler-Hayut, Kiril Kniazev, Cathy Melamed-Bessudo, Avraham A. Levy
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	homologous recombination DNA repair CRISPR Cas9 chromatin Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/8754864f9c0740b3adac1f46c3886a64
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spelling	oai:doaj.org-article:8754864f9c0740b3adac1f46c3886a642021-11-25T17:53:24ZTargeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin10.3390/ijms2222120961422-00671661-6596https://doaj.org/article/8754864f9c0740b3adac1f46c3886a642021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12096https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Homologous 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 <i>Arabidopsis thaliana</i>. 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.Shdema Filler-HayutKiril KniazevCathy Melamed-BessudoAvraham A. LevyMDPI AGarticlehomologous recombinationDNA repairCRISPR Cas9chromatinBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12096, p 12096 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	homologous recombination DNA repair CRISPR Cas9 chromatin Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	homologous recombination DNA repair CRISPR Cas9 chromatin Biology (General) QH301-705.5 Chemistry QD1-999 Shdema Filler-Hayut Kiril Kniazev Cathy Melamed-Bessudo Avraham A. Levy Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
description	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 <i>Arabidopsis thaliana</i>. 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.
format	article
author	Shdema Filler-Hayut Kiril Kniazev Cathy Melamed-Bessudo Avraham A. Levy
author_facet	Shdema Filler-Hayut Kiril Kniazev Cathy Melamed-Bessudo Avraham A. Levy
author_sort	Shdema Filler-Hayut
title	Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
title_short	Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
title_full	Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
title_fullStr	Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
title_full_unstemmed	Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin
title_sort	targeted inter-homologs recombination in arabidopsis euchromatin and heterochromatin
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/8754864f9c0740b3adac1f46c3886a64
work_keys_str_mv	AT shdemafillerhayut targetedinterhomologsrecombinationinarabidopsiseuchromatinandheterochromatin AT kirilkniazev targetedinterhomologsrecombinationinarabidopsiseuchromatinandheterochromatin AT cathymelamedbessudo targetedinterhomologsrecombinationinarabidopsiseuchromatinandheterochromatin AT avrahamalevy targetedinterhomologsrecombinationinarabidopsiseuchromatinandheterochromatin
_version_	1718411872709902336

Targeted Inter-Homologs Recombination in Arabidopsis Euchromatin and Heterochromatin

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