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...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2018-03-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/31677 |
| _version_ | 1818024998564003840 |
|---|---|
| author | Frederic Strobl Anita Anderl Ernst HK Stelzer |
| author_facet | Frederic Strobl Anita Anderl Ernst HK Stelzer |
| author_sort | Frederic Strobl |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T04:09:07Z |
| format | Article |
| id | doaj.art-377f5d21392643f3963071605b53945b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T04:09:07Z |
| publishDate | 2018-03-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-377f5d21392643f3963071605b53945b2022-12-22T02:02:47ZengeLife Sciences Publications LtdeLife2050-084X2018-03-01710.7554/eLife.31677A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous linesFrederic Strobl0Anita Anderl1Ernst HK Stelzer2https://orcid.org/0000-0003-1545-0736Physical Biology, BMLS, CEF-MC, Goethe Universität, Frankfurt am Main, GermanyPhysical Biology, BMLS, CEF-MC, Goethe Universität, Frankfurt am Main, GermanyPhysical Biology, BMLS, CEF-MC, Goethe Universität, Frankfurt am Main, GermanyDiploid 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.https://elifesciences.org/articles/31677Tribolium castaneumgenotypingtransgenesisCre-Lox systemlong-term live imaginglight sheet-based fluorescence microscopy |
| spellingShingle | Frederic Strobl Anita Anderl Ernst HK Stelzer A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines eLife Tribolium castaneum genotyping transgenesis Cre-Lox system long-term live imaging light sheet-based fluorescence microscopy |
| title | A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| title_full | A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| title_fullStr | A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| title_full_unstemmed | A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| title_short | A universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| title_sort | universal vector concept for a direct genotyping of transgenic organisms and a systematic creation of homozygous lines |
| topic | Tribolium castaneum genotyping transgenesis Cre-Lox system long-term live imaging light sheet-based fluorescence microscopy |
| url | https://elifesciences.org/articles/31677 |
| work_keys_str_mv | AT fredericstrobl auniversalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines AT anitaanderl auniversalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines AT ernsthkstelzer auniversalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines AT fredericstrobl universalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines AT anitaanderl universalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines AT ernsthkstelzer universalvectorconceptforadirectgenotypingoftransgenicorganismsandasystematiccreationofhomozygouslines |