A conditional tissue-specific transgene expression system using inducible GAL4

Proceedings of the National Academy of Sciences of the United States of America - Tập 98 Số 22 - Trang 12596-12601 - 2001
Thomas Osterwalder1, Kenneth S. Yoon2, Benjamin H. White3,2, Haig Keshishian2
1Department of Molecular, Cellular, and Developmental Biology, and Pharmacology Department, Yale University, P. O. Box 208103, New Haven, CT 06511, USA.
2Department of Molecular, Cellular, and Developmental Biology, and Pharmacology Department, Yale University, P. O. Box 208103, New Haven, CT 06511
3Department of Molecular, Cellular and Developmental Biology and

Tóm tắt

In Drosophila , the most widely used system for generating spatially restricted transgene expression is based on the yeast GAL4 protein and its target upstream activating sequence (UAS). To permit temporal as well as spatial control over UAS-transgene expression, we have explored the use of a conditional RU486-dependent GAL4 protein (GeneSwitch) in Drosophila . By using cloned promoter fragments of the embryonic lethal abnormal vision gene or the myosin heavy chain gene, we have expressed GeneSwitch specifically in neurons or muscles and show that its transcriptional activity within the target tissues depends on the presence of the activator RU486 (mifepristone). We used available UAS-reporter lines to demonstrate RU486-dependent tissue-specific transgene expression in larvae. Reporter protein expression could be detected 5 h after systemic application of RU486 by either feeding or “larval bathing.” Transgene expression levels were dose-dependent on RU486 concentration in larval food, with low background expression in the absence of RU486. By using genetically altered ion channels as reporters, we were able to change the physiological properties of larval bodywall muscles in an RU486-dependent fashion. We demonstrate here the applicability of GeneSwitch for conditional tissue-specific expression in Drosophila , and we provide tools to control pre- and postsynaptic expression of transgenes at the larval neuromuscular junction during postembryonic life.

Từ khóa


Tài liệu tham khảo

10.1242/dev.118.2.401

10.1101/gad.8.15.1787

10.1139/g96-023

10.1016/0959-4388(95)80061-1

10.1006/meth.1998.0592

10.1016/S0896-6273(00)00154-9

10.1073/pnas.94.10.5131

10.1016/0896-6273(95)90290-2

10.1016/0896-6273(94)90045-0

10.1016/S0896-6273(00)80154-3

10.1073/pnas.94.12.6255

10.1126/science.277.5327.825

10.1242/dev.125.12.2193

10.1016/S0378-1119(01)00447-4

10.1007/s004380050770

10.1016/0958-1669(94)90067-1

10.1126/science.7792603

10.1073/pnas.93.22.12418

10.1242/dev.125.6.1049

10.1073/pnas.90.5.1657

M M Burcin, B W O'Malley, S Y Tsai Front Biosci 3, c1–c7 (1998).

10.1242/dev.127.3.573

10.1073/pnas.96.2.355

10.1073/pnas.221303998

10.1126/science.6289436

10.1038/227680a0

10.1073/pnas.76.9.4350

10.1002/neu.480240606

10.1046/j.1471-4159.2000.0741941.x

10.1146/annurev.ento.45.1.151

10.1074/jbc.273.52.34970

10.1038/78531

L M Riddiford The Embryonic Development of Drosophila melanogaster (Cold Spring Harbor Lab. Press, Plainview, NY, 1997).

V Budnik, L S Gramates Neuromuscular Junctions in Drosophila, 1999).

10.1016/S0896-6273(01)00415-9

Thông tin
Thông tin xuất bản

Proceedings of the National Academy of Sciences of the United States of America

Tập 98 Số 22

12596-12601

Thông tin tác giả