Gene-specific cell labeling using MiMIC transposons.
Gene-specific cell labeling using MiMIC transposons.
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DOI：
10.1093/nar/gkv113
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发表时间：
2015-04-30
影响因子：
14.9
通讯作者：
Dierick HA
中科院分区：
文献类型：
--
作者：
Gnerer JP;Venken KJ;Dierick HA
GAL 4/UAS、莱克萨/LexAop和QF/QUAS等二元表达系统通过允许时空操纵基因功能以及细胞和神经回路功能，极大地增强了果蝇作为模式生物的能力。这些异源转录因子的组织特异性表达依赖于随机转座子整合在驱动嵌入转座子中的二元转录因子的增强子或启动子附近。或者，基因特异性启动子元件直接融合到转座子内的二元因子，然后随机或位点特异性整合。然而，这样的插入并不一致地概括内源性表达。我们使用Minos介导的整合盒（MiMIC）转座子将宿主基因座转换为可靠的基因特异性二元效应子。MiMIC转座子允许重组酶介导的盒交换以修改转座子内容。我们开发了新的交换盒，将编码内含子MiMIC插入转换为基因特异性二元因子蛋白陷阱。此外，我们扩展了一组可用于非编码内含子MiMIC插入的二元因子交换盒。我们发现，在同一位点的不同插入的二元因子转换具有不可区分的表达模式，这表明它们可靠地反映了内源基因表达。我们通过剖析果蝇5-羟色胺受体基因家族的细胞表达模式，展示了这些新工具的有效性和广泛适用性。
Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family.
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影响因子：
3.7
作者：
Becnel J;Johnson O;Luo J;Nässel DR;Nichols CD
通讯作者：
Nichols CD
DOI：
10.1083/jcb.150.2.f83
发表时间：
2000-07-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Brody T;Cravchik A
通讯作者：
Cravchik A
影响因子：
9.8
作者：
Fayyazuddin A;Zaheer MA;Hiesinger PR;Bellen HJ
通讯作者：
Bellen HJ
影响因子：
14.9
作者：
Grumbling G;Strelets V
通讯作者：
Strelets V
影响因子：
9.2
作者：
Chan, Chih-Chiang;Scoggin, Shane;Wang, Dong;Cherry, Smita;Dembo, Todd;Greenberg, Ben;Jin, Eugene Jennifer;Kuey, Cansu;Lopez, Antonio;Mehta, Sunil Q.;Perkins, Theodore J.;Brankatschk, Marko;Rothenfluh, Adrian;Buszczak, Michael;Hiesinger, P. Robin
通讯作者：
Hiesinger, P. Robin