Abstract
Artificial minichromosomes are highly desirable tools for basic research, breeding, and biotechnology purposes. We present an option to generate plant artificial minichromosomes via de novo engineering of plant centromeres in Arabidopsis thaliana by targeting kinetochore proteins to tandem repeat arrays at non-centromeric positions. We employed the bacterial lactose repressor/lactose operator system to guide derivatives of the centromeric histone H3 variant cenH3 to LacO operator sequences. Tethering of cenH3 to non-centromeric loci led to de novo assembly of kinetochore proteins and to dicentric carrier chromosomes which potentially form anaphase bridges. This approach will be further developed and may contribute to generating minichromosomes from preselected genomic regions, potentially even in a diploid background.
Original language | English (US) |
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Pages (from-to) | 233-241 |
Number of pages | 9 |
Journal | Chromosoma |
Volume | 122 |
Issue number | 3 |
DOIs | |
State | Published - Jun 2013 |
Externally published | Yes |
Bibliographical note
Funding Information:Acknowledgments We would like to thank Eric Lam for providing line EL702C, Markus Kuhlmann for the helpful suggestions, Andreas Finke for the T-DNA binary vectors, Twan Rutten for the help with confocal microscopy, Minoru Murata and Kiyotaka Nagaki for providing the AtMIS12 antibody, and Katrin Kumke, Christa Fricke, Inge Glaser, and Beate Kamm for the excellent technical assistance. This project was partly supported by the Leibniz Association (WGL, “Pakt für Forschung und Innovation”) and by the IPK.
Keywords
- De novo centromere formation
- Kinetochore proteins
- Lac operator system
- Lac repressor
- Technical advance
- cenH3 targeting
ASJC Scopus subject areas
- Genetics
- Genetics(clinical)