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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| Format: | Online |
| Language: | English |
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eLife Sciences Publications, Ltd
2015
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4379497/ |
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pubmed-4379497 |
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oai_dc |
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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 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. 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. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| 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 |
| spellingShingle |
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 |
| 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 |
| title |
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_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_sort |
library of mimics allows tagging of genes and reversible, spatial and temporal knockdown of proteins in drosophila |
| 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. |
| publisher |
eLife Sciences Publications, Ltd |
| publishDate |
2015 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4379497/ |
| _version_ |
1613205550663729152 |