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PLoS One. 2013 Dec 26;8(12):e85383. doi: 10.1371/journal.pone.0085383. eCollection 2013.

Epigenetic alterations at genomic loci modified by gene targeting in Arabidopsis thaliana.

Author information

1
Department of Plant Biology, University of Geneva, Geneva, Switzerland ; Department of Plant Sciences, The Weizmann Institute of Science, Rehovot, Israel.
2
Department of Plant Sciences, The Weizmann Institute of Science, Rehovot, Israel.
3
Department of Molecular and Developmental Genetics, Institute of Biology, Leiden University, Leiden, The Netherlands.

Abstract

Gene Targeting (GT) is the integration of an introduced vector into a specific chromosomal site, via homologous recombination. It is considered an effective tool for precise genome editing, with far-reaching implications in biological research and biotechnology, and is widely used in mice, with the potential of becoming routine in many species. Nevertheless, the epigenetic status of the targeted allele remains largely unexplored. Using GT-modified lines of the model plant Arabidopsis thaliana, we show that the DNA methylation profile of the targeted locus is changed following GT. This effect is non-directional as methylation can be either completely lost, maintained with minor alterations or show instability in the generations subsequent to GT. As DNA methylation is known to be involved in several cellular processes, GT-related alterations may result in unexpected or even unnoticed perturbations. Our analysis shows that GT may be used as a new tool for generating epialleles, for example, to study the role of gene body methylation. In addition, the analysis of DNA methylation at the targeted locus may be utilized to investigate the mechanism of GT, many aspects of which are still unknown.

PMID:
24386472
PMCID:
PMC3873452
DOI:
10.1371/journal.pone.0085383
[Indexed for MEDLINE]
Free PMC Article

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