Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, UK.
Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, USA.
Sci Rep. 2022 Mar 9;12(1):3835. doi: 10.1038/s41598-022-07852-7.
The existence of three independent binary systems for conditional gene expression (Gal4/UAS; LexA/LexAop; QF/QUAS) has greatly expanded versatile genetic analyses in the Drosophila melanogaster; however, the experimental application of these tools is limited by the need to generate multiple collections of noninterchangeable transgenic fly strains for each inducible gene expression system. To address this practical limitation, we developed a modular vector that contains the regulatory elements from all three binary systems, enabling Gal4-, LexA- or QF-dependent expression of transgenes. Our methods also incorporate DNA elements that facilitate independent site-specific recombination and elimination of regulatory UAS, LexAop or QUAS modules with spatial and temporal control, thus offering unprecedented possibilities and logistical advantages for in vivo genetic modulation and efficient interconversion of overexpression transgenic fly lines.
三种独立的条件基因表达二元系统（Gal4/UAS；LexA/LexAop；QF/QUAS）的存在极大地扩展了黑腹果蝇中的多功能遗传分析；然而，这些工具的实验应用受到需要为每个诱导基因表达系统生成多组不可互换的转基因果蝇品系的限制。为了解决这个实际限制，我们开发了一种模块化载体，其中包含所有三个二元系统的调控元件，使转基因能够依赖 Gal4、LexA 或 QF 进行表达。我们的方法还包含有助于独立的位点特异性重组和具有时空控制的 UAS、LexAop 或 QUAS 调控模块的消除的 DNA 元件，从而为体内遗传调节和过表达转基因果蝇品系的高效相互转换提供了前所未有的可能性和逻辑优势。
