Coordination of Pickpocket ion channel delivery and dendrite growth in Drosophila sensory neurons

Mitchell, Josephine W.; Midillioglu, Ipek; Wang, Bei; Han, Chun; Wildonger, Jill

doi:10.1101/2022.05.19.492359

Cited by 1 publication

(2 citation statements)

References 92 publications

(159 reference statements)

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“…Historically, full-length FPs have been widely used in various applications. More recently, the split GFP1-10/11 system has emerged as a valuable tool for live-cell protein imaging, especially in tissue imaging, due to its binary nature, which facilitates cell-type-specific labeling [5][6][7][8][9]. In this system, the super-folder GFP sequence is divided between b-strand 10 and 11 (GFP1-10 and GFP11) [10].…”

Section: Introductionmentioning

confidence: 99%

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Cell-type-specific Labeling of Endogenous Proteins Using the Split GFP System inDrosophila

Inal,

Banzai,

Kamiyama

et al. 2024

Preprint

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Accurate identification of the locations of endogenous proteins is crucial for understanding their functions in tissues and cells. However, achieving precise cell-type-specific labeling of these proteins has been challenging in vivo. A notable solution to this challenge is the self-complementing split green fluorescent protein (GFP1-10/11) system. In this paper, we present a detailed protocol for labeling endogenous proteins in a cell-type-specific manner using the GFP1-10/11 system in fruit flies. This approach depends on the automatic reconstitution of the GFP1-10 and GFP11 fragments, creating a fluorescence signal. We insert the GFP11 fragment into a specific genomic locus while expressing its counterpart, GFP1-10, through an available Gal4 driver line. The unique aspect of this system is that neither GFP1-10 nor GFP11 alone emits fluorescence, enabling the precise detection of protein localization only in Gal4-positive cells. We illustrate this technique using the adhesion molecule gene teneurin-m (Ten-m) as a model, highlighting the generation and validation of GFP11 protein trap lines via Minos-mediated integration cassette (MiMIC) insertion. Furthermore, we demonstrate the cell-type-specific labeling of Ten-m proteins in the larval brains of fruit flies. This method significantly enhances our ability to image endogenous protein localization patterns in a cell-type-specific manner and is adaptable to various model organisms beyond fruit flies.

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Section: Introductionmentioning

confidence: 99%

“…Furthermore, due to its binary nature reconstitution can be achieved in a cell-specific manner [5][6][7][8][9]. In this system, the super-folder GFP sequence is divided between b-strand 10 and 11 (GFP1-10 and GFP11) [10].…”

Section: Introductionmentioning

confidence: 99%