Expansion of the gateway multisite recombination cloning toolkit.
Precise manipulation of transgene expression in genetic model organisms has led to advances in understanding fundamental mechanisms of development, physiology, and genetic disease. Transgene construction is, however, a precondition of transgene expression, and often limits the rate of experimental p...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3799639?pdf=render |
| _version_ | 1818580505328615424 |
|---|---|
| author | Harold K Shearin Alisa R Dvarishkis Craig D Kozeluh R Steven Stowers |
| author_facet | Harold K Shearin Alisa R Dvarishkis Craig D Kozeluh R Steven Stowers |
| author_sort | Harold K Shearin |
| collection | DOAJ |
| description | Precise manipulation of transgene expression in genetic model organisms has led to advances in understanding fundamental mechanisms of development, physiology, and genetic disease. Transgene construction is, however, a precondition of transgene expression, and often limits the rate of experimental progress. Here we report an expansion of the modular Gateway MultiSite recombination-cloning platform for high efficiency transgene assembly. The expansion includes two additional destination vectors and entry clones for the LexA binary transcription system, among others. These new tools enhance the expression levels possible with Gateway MultiSite generated transgenes and make possible the generation of LexA drivers and reporters with Gateway MultiSite cloning. In vivo data from transgenic Drosophila functionally validating each novel component are presented and include neuronal LexA drivers, LexAop2 red and green fluorescent synaptic vesicle reporters, TDC2 and TRH LexA, GAL4, and QF drivers, and LexAop2, UAS, and QUAS channelrhodopsin2 T159C reporters. |
| first_indexed | 2024-12-16T07:18:40Z |
| format | Article |
| id | doaj.art-ad88b35890f34a97863dc35520f65cfd |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-16T07:18:40Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ad88b35890f34a97863dc35520f65cfd2022-12-21T22:39:43ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01810e7772410.1371/journal.pone.0077724Expansion of the gateway multisite recombination cloning toolkit.Harold K ShearinAlisa R DvarishkisCraig D KozeluhR Steven StowersPrecise manipulation of transgene expression in genetic model organisms has led to advances in understanding fundamental mechanisms of development, physiology, and genetic disease. Transgene construction is, however, a precondition of transgene expression, and often limits the rate of experimental progress. Here we report an expansion of the modular Gateway MultiSite recombination-cloning platform for high efficiency transgene assembly. The expansion includes two additional destination vectors and entry clones for the LexA binary transcription system, among others. These new tools enhance the expression levels possible with Gateway MultiSite generated transgenes and make possible the generation of LexA drivers and reporters with Gateway MultiSite cloning. In vivo data from transgenic Drosophila functionally validating each novel component are presented and include neuronal LexA drivers, LexAop2 red and green fluorescent synaptic vesicle reporters, TDC2 and TRH LexA, GAL4, and QF drivers, and LexAop2, UAS, and QUAS channelrhodopsin2 T159C reporters.http://europepmc.org/articles/PMC3799639?pdf=render |
| spellingShingle | Harold K Shearin Alisa R Dvarishkis Craig D Kozeluh R Steven Stowers Expansion of the gateway multisite recombination cloning toolkit. PLoS ONE |
| title | Expansion of the gateway multisite recombination cloning toolkit. |
| title_full | Expansion of the gateway multisite recombination cloning toolkit. |
| title_fullStr | Expansion of the gateway multisite recombination cloning toolkit. |
| title_full_unstemmed | Expansion of the gateway multisite recombination cloning toolkit. |
| title_short | Expansion of the gateway multisite recombination cloning toolkit. |
| title_sort | expansion of the gateway multisite recombination cloning toolkit |
| url | http://europepmc.org/articles/PMC3799639?pdf=render |
| work_keys_str_mv | AT haroldkshearin expansionofthegatewaymultisiterecombinationcloningtoolkit AT alisardvarishkis expansionofthegatewaymultisiterecombinationcloningtoolkit AT craigdkozeluh expansionofthegatewaymultisiterecombinationcloningtoolkit AT rstevenstowers expansionofthegatewaymultisiterecombinationcloningtoolkit |