PUBLICATION
A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines
- Authors
- Strobl, F., Anderl, A., Stelzer, E.H.
- ID
- ZDB-PUB-180316-9
- Date
- 2018
- Source
- eLIFE 7: (Journal)
- Registered Authors
- Keywords
- Cre-Lox system, Tribolium castaneum, chromosomes, developmental biology, genes, genotyping, light sheet-based fluorescence microscopy, long-term live imaging, stem cells, transgenesis
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics*
- Diploidy*
- Genetic Vectors
- Genotype
- Hemizygote
- Homozygote
- Insecta/genetics
- Integrases/genetics
- Plants/genetics
- Plants, Genetically Modified/genetics*
- Recombination, Genetic/genetics*
- Rodentia/genetics
- Tribolium/genetics
- Zebrafish/genetics
- PubMed
- 29543587 Full text @ Elife
Citation
Strobl, F., Anderl, A., Stelzer, E.H. (2018) A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines. eLIFE. 7.
Abstract
Diploid transgenic organisms are either hemi- or homozygous. Genetic assays are, therefore, required to identify the genotype. Our AGameOfClones vector concept uses two clearly distinguishable transformation markers embedded in interweaved, but incompatible Lox site pairs. Cre-mediated recombination leads to hemizygous individuals that carry only one marker. In the following generation, heterozygous descendants are identified by the presence of both markers and produce homozygous progeny that are selected by the lack of one marker. We prove our concept in Tribolium castaneum by systematically creating multiple functional homozygous transgenic lines suitable for long-term fluorescence live imaging. Our approach saves resources and simplifies transgenic organism handling. Since the concept relies on the universal Cre-Lox system, it is expected to work in all diploid model organisms, for example, insects, zebrafish, rodents and plants. With appropriate adaptions, it can be used in knock-out assays to preselect homozygous individuals and thus minimize the number of wasted animals.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping