A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila
Sonal Nagarkar-Jaiswal,
Pei-Tseng Lee,
Megan E Campbell,
Kuchuan Chen,
Stephanie Anguiano-Zarate,
Manuel Cantu Gutierrez,
Theodore Busby,
Wen-Wen Lin,
Yuchun He,
Karen L Schulze,
Benjamin W Booth,
Martha Evans-Holm,
Koen JT Venken,
Robert W Levis,
Allan C Spradling,
Roger A Hoskins,
Hugo J Bellen
Affiliations
Sonal Nagarkar-Jaiswal
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Pei-Tseng Lee
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Megan E Campbell
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Kuchuan Chen
Program in Developmental Biology, Baylor College of Medicine, Houston, United States
Stephanie Anguiano-Zarate
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Manuel Cantu Gutierrez
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Theodore Busby
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Wen-Wen Lin
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
Yuchun He
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
Karen L Schulze
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
Benjamin W Booth
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, United States
Martha Evans-Holm
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, United States
Koen JT Venken
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, United States
Robert W Levis
Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
Allan C Spradling
Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
Roger A Hoskins
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, United States
Hugo J Bellen
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Program in Developmental Biology, Baylor College of Medicine, Houston, United States; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, United States; Department of Neuroscience, Baylor College of Medicine, Houston, United States
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.
