A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines
- 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.
Author: | Frederic StroblORCiDGND, Anita Anderl, Ernst H. K. StelzerORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-463769 |
DOI: | https://doi.org/10.7554/eLife.31677 |
ISSN: | 2050-084X |
Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/29543587 |
Parent Title (English): | eLife |
Publisher: | eLife Sciences Publications |
Place of publication: | Cambridge |
Contributor(s): | Bruce Edgar |
Document Type: | Article |
Language: | English |
Year of Completion: | 2018 |
Date of first Publication: | 2018/03/15 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2018/05/15 |
Tag: | Chromosomes and gene expression; Cre-lox system; Developmental biology and stem cells; Genotyping; Light sheet-based fluorescence microscopy; Long-term live imaging; Tools and resources; Transgenesis; Tribolium castaneum |
Volume: | 7 |
Issue: | e31677 |
Page Number: | 21 |
First Page: | 1 |
Last Page: | 21 |
Note: | 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. |
HeBIS-PPN: | 433868910 |
Institutes: | Biowissenschaften / Biowissenschaften |
Exzellenzcluster / Exzellenzcluster Makromolekulare Komplexe | |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
Sammlungen: | Universitätspublikationen |
Sammlung Biologie / Sondersammelgebiets-Volltexte | |
Open-Access-Publikationsfonds: | Biowissenschaften |
Licence (German): | Creative Commons - Namensnennung 4.0 |