Journal
ELIFE
Volume 4, Issue -, Pages -Publisher
ELIFE SCIENCES PUBLICATIONS LTD
DOI: 10.7554/eLife.05338
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Funding
- National Institute of General Medical Sciences (NIGMS)
- National Institute of Child Health and Human Development (NICHD) [R01GM067858]
- March of Dimes Foundation [3R01GM067858-11S1, 1-FY14-315]
- Cancer Prevention and Research Institute of Texas (CPRIT)
- National Human Genome Research Institute (NHGRI)
- National Institute of General Medical Sciences (NIGMS) [3R01GM067858-09S1]
- Baylor College of Medicine
- National Institutes of Health (NIH) [1R21GM110190]
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Here, we document a collection of similar to 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.
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