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A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be image...

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Autores principales: Nagarkar-Jaiswal, Sonal, Lee, Pei-Tseng, Campbell, Megan E, Chen, Kuchuan, Anguiano-Zarate, Stephanie, Cantu Gutierrez, Manuel, Busby, Theodore, Lin, Wen-Wen, He, Yuchun, Schulze, Karen L, Booth, Benjamin W, Evans-Holm, Martha, Venken, Koen JT, Levis, Robert W, Spradling, Allan C, Hoskins, Roger A, Bellen, Hugo J
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4379497/
https://www.ncbi.nlm.nih.gov/pubmed/25824290
http://dx.doi.org/10.7554/eLife.05338
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author Nagarkar-Jaiswal, Sonal
Lee, Pei-Tseng
Campbell, Megan E
Chen, Kuchuan
Anguiano-Zarate, Stephanie
Cantu Gutierrez, Manuel
Busby, Theodore
Lin, Wen-Wen
He, Yuchun
Schulze, Karen L
Booth, Benjamin W
Evans-Holm, Martha
Venken, Koen JT
Levis, Robert W
Spradling, Allan C
Hoskins, Roger A
Bellen, Hugo J
author_facet Nagarkar-Jaiswal, Sonal
Lee, Pei-Tseng
Campbell, Megan E
Chen, Kuchuan
Anguiano-Zarate, Stephanie
Cantu Gutierrez, Manuel
Busby, Theodore
Lin, Wen-Wen
He, Yuchun
Schulze, Karen L
Booth, Benjamin W
Evans-Holm, Martha
Venken, Koen JT
Levis, Robert W
Spradling, Allan C
Hoskins, Roger A
Bellen, Hugo J
author_sort Nagarkar-Jaiswal, Sonal
collection PubMed
description Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be imaged in unfixed tissues. Moreover, the tagged mRNAs can be knocked down by RNAi against GFP (iGFPi), and the tagged proteins can be efficiently knocked down by deGradFP technology. The phenotypes associated with RNA and protein knockdown typically correspond to severe loss of function or null mutant phenotypes. Finally, we demonstrate reversible, spatial, and temporal knockdown of tagged proteins in larvae and adult flies. This new strategy and collection of strains allows unprecedented in vivo manipulations in flies for many genes. These strategies will likely extend to vertebrates. DOI: http://dx.doi.org/10.7554/eLife.05338.001
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spelling pubmed-43794972015-04-02 A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila Nagarkar-Jaiswal, Sonal Lee, Pei-Tseng Campbell, Megan E Chen, Kuchuan Anguiano-Zarate, Stephanie Cantu Gutierrez, Manuel Busby, Theodore Lin, Wen-Wen He, Yuchun Schulze, Karen L Booth, Benjamin W Evans-Holm, Martha Venken, Koen JT Levis, Robert W Spradling, Allan C Hoskins, Roger A Bellen, Hugo J eLife Cell Biology Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be imaged in unfixed tissues. Moreover, the tagged mRNAs can be knocked down by RNAi against GFP (iGFPi), and the tagged proteins can be efficiently knocked down by deGradFP technology. The phenotypes associated with RNA and protein knockdown typically correspond to severe loss of function or null mutant phenotypes. Finally, we demonstrate reversible, spatial, and temporal knockdown of tagged proteins in larvae and adult flies. This new strategy and collection of strains allows unprecedented in vivo manipulations in flies for many genes. These strategies will likely extend to vertebrates. DOI: http://dx.doi.org/10.7554/eLife.05338.001 eLife Sciences Publications, Ltd 2015-03-31 /pmc/articles/PMC4379497/ /pubmed/25824290 http://dx.doi.org/10.7554/eLife.05338 Text en © 2015, Nagarkar-Jaiswal et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Nagarkar-Jaiswal, Sonal
Lee, Pei-Tseng
Campbell, Megan E
Chen, Kuchuan
Anguiano-Zarate, Stephanie
Cantu Gutierrez, Manuel
Busby, Theodore
Lin, Wen-Wen
He, Yuchun
Schulze, Karen L
Booth, Benjamin W
Evans-Holm, Martha
Venken, Koen JT
Levis, Robert W
Spradling, Allan C
Hoskins, Roger A
Bellen, Hugo J
A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title_full A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title_fullStr A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title_full_unstemmed A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title_short A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
title_sort library of mimics allows tagging of genes and reversible, spatial and temporal knockdown of proteins in drosophila
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4379497/
https://www.ncbi.nlm.nih.gov/pubmed/25824290
http://dx.doi.org/10.7554/eLife.05338
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