Expanding the Drosophila toolkit for dual control of gene expression

被引:3
|
作者
Zirin, Jonathan [1 ]
Jusiak, Barbara [2 ]
Lopes, Raphael [1 ]
Ewen-Campen, Benjamin [1 ]
Bosch, Justin A. [1 ]
Risbeck, Alexandria [1 ]
Forman, Corey [1 ]
Villalta, Christians [1 ]
Hu, Yanhui [1 ]
Perrimon, Norbert [1 ,3 ]
机构
[1] Harvard Med Sch, Dept Genet, Boston, MA 02115 USA
[2] Univ Calif Irvine, Dept Physiol & Biophys, Irvine, CA USA
[3] Howard Hughes Med Inst, Boston, MA 02115 USA
来源
ELIFE | 2024年 / 12卷
关键词
LexA; QF; Drosophila; GAL4; CRISPR; inter-organ communication; D; melanogaster; Q-SYSTEM; TRANSGENIC RNAI; DECAPENTAPLEGIC GENE; RESOURCE; INSERTION;
D O I
10.7554/eLife.94073
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The ability to independently control gene expression in two different tissues in the same animal is emerging as a major need, especially in the context of inter-organ communication studies. This type of study is made possible by technologies combining the GAL4/UAS and a second binary expression system such as the LexA system or QF system. Here, we describe a resource of reagents that facilitate combined use of the GAL4/UAS and a second binary system in various Drosophila tissues. Focusing on genes with well-characterized GAL4 expression patterns, we generated a set of more than 40 LexA-GAD and QF2 insertions by CRISPR knock-in and verified their tissue specificity in larvae. We also built constructs that encode QF2 and LexA-GAD transcription factors in a single vector. Following successful integration of this construct into the fly genome, FLP/FRT recombination is used to isolate fly lines that express only QF2 or LexA-GAD. Finally, using new compatible shRNA vectors, we evaluated both LexA and QF systems for in vivo gene knockdown and are generating a library of such RNAi fly lines as a community resource. Together, these LexA and QF system vectors and fly lines will provide a new set of tools for researchers who need to activate or repress two different genes in an orthogonal manner in the same animal.
引用
收藏
页数:19
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