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

Abstract

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.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [2];  [1];  [3];  [3];  [2];  [4]
  1. Baylor College of Medicine, Houston, TX (United States)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. Howard Hughes Medical Institute, Baltimore, MD (United States)
  4. Baylor College of Medicine, Houston, TX (United States); Texas Children's Hospital, Houston, TX (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1190953
Grant/Contract Number:
R01GM067858; 3R01GM067858-11S1; 3R01GM067858-09S1; McNair Startup Funds; #1-FY14-315; 1R21GM110190
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 4; Journal Issue: 3; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English

Citation Formats

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 J.T., Levis, Robert W., Spradling, Allan C., Hoskins, Roger A., and Bellen, Hugo J.. A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila. United States: N. p., 2015. Web. doi:10.7554/eLife.05338.
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 J.T., 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. United States. doi:10.7554/eLife.05338.
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 J.T., Levis, Robert W., Spradling, Allan C., Hoskins, Roger A., and Bellen, Hugo J.. Tue . "A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila". United States. doi:10.7554/eLife.05338. https://www.osti.gov/servlets/purl/1190953.
@article{osti_1190953,
title = {A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila},
author = {Nagarkar-Jaiswal, Sonal and Lee, Pei-Tseng and Campbell, Megan E. and Chen, Kuchuan and Anguiano-Zarate, Stephanie and Cantu Gutierrez, Manuel and Busby, Theodore and Lin, Wen-Wen and He, Yuchun and Schulze, Karen L. and Booth, Benjamin W. and Evans-Holm, Martha and Venken, Koen J.T. and Levis, Robert W. and Spradling, Allan C. and Hoskins, Roger A. and Bellen, Hugo J.},
abstractNote = {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 = {10.7554/eLife.05338},
journal = {eLife},
number = 3,
volume = 4,
place = {United States},
year = {Tue Mar 31 00:00:00 EDT 2015},
month = {Tue Mar 31 00:00:00 EDT 2015}
}

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