TY  - JOUR
A1  - Strobl, Frederic
A1  - Anderl, Anita
A1  - Stelzer, Ernst H. K.
T1  - A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines
T2  - eLife
N2  - 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.
KW  - Tools and resources
KW  - Developmental biology and stem cells
KW  - Chromosomes and gene expression
KW  - Tribolium castaneum
KW  - Genotyping
KW  - Transgenesis
KW  - Cre-lox system
KW  - Long-term live imaging
KW  - Light sheet-based fluorescence microscopy
Y1  - 2018
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/46376
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-463769
SN  - 2050-084X
N1  - Copyright Strobl et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
VL  - 7
IS  - e31677
SP  - 1
EP  - 21
PB  - eLife Sciences Publications
CY  - Cambridge
ER  -