Marker-Free Transgenic Plants through Genetically Programmed Auto-Excision

We present here a vector system to obtain homozygous marker-free transgenic plants without the need of extra handling and within the same time frame as compared to transformation methods in which the marker is not removed. By introducing a germline-specific auto-excision vector containing a cre reco...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 2007-12, Vol.145 (4), p.1220-1231
Main Authors: Verweire, Dimitri, Verleyen, Kristof, De Buck, Sylvie, Claeys, Martine, Angenon, Geert
Format: Article
Language:English
Subjects:
Alleles
Application programming interfaces
Arabidopsis - genetics
Arabidopsis - metabolism
Arabidopsis Proteins - genetics
Arabidopsis thaliana
Breakthrough Technologies
Cyclins - genetics
DNA Cleavage
DNA, Bacterial
Genetic Engineering
Genetic loci
Genetic Markers
Genetic vectors
Germ cells
Germ Cells - metabolism
MADS Domain Proteins - genetics
Plants
Plants, Genetically Modified - metabolism
Polymerase chain reaction
Promoter Regions, Genetic
Segregation
Transgenes
Transgenic plants
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Summary:We present here a vector system to obtain homozygous marker-free transgenic plants without the need of extra handling and within the same time frame as compared to transformation methods in which the marker is not removed. By introducing a germline-specific auto-excision vector containing a cre recombinase gene under the control of a germline-specific promoter, transgenic plants become genetically programmed to lose the marker when its presence is no longer required (i.e. after the initial selection of primary transformants). Using promoters with different germline functionality, two modules of this genetic program were developed. In the first module, the promoter, placed upstream of the cre gene, confers CRE functionality in both the male and the female germline or in the common germline (e.g. floral meristem cells). In the second module, a promoter conferring single germline-specific CRE functionality was introduced upstream of the cre gene. Promoter sequences used in this work are derived from the APETALA1 and SOLO DANCERS genes from Arabidopsis (Arabidopsis thaliana) Columbia-0 conferring common germline and single germline functionality, respectively. Introduction of the genetic program did not reduce transformation efficiency. Marker-free homozygous progeny plants were efficiently obtained, regardless of which promoter was used. In addition, simplification of complex transgene loci was observed.
ISSN:0032-0889
1532-2548
1532-2548
DOI:10.1104/pp.107.106526