Fly-FUCCI: A versatile tool for studying cell proliferation in complex tissues.

Area of Science:

• Cell Biology
• Developmental Biology
• Genetics

Background:

• The FUCCI (fluorescent ubiquitination-based cell cycle indicator) system is a powerful tool for real-time cell cycle analysis.
• Adapting such systems for model organisms like Drosophila is crucial for understanding fundamental biological processes.

Purpose of the Study:

• To develop and validate a Drosophila-specific FUCCI system (Fly-FUCCI) for distinguishing G1, S, and G2 phases of interphase.
• To create a versatile toolkit of transgenic Drosophila lines for broad experimental applications.

Main Methods:

• Engineered Fly-FUCCI probes using fluorochrome-tagged degrons from Cyclin B and E2F1 proteins.
• Utilized ubiquitin E3-ligases APC/C and CRL4(Cdt2) for targeted protein degradation.
• Generated transgenic Drosophila lines expressing Fly-FUCCI probes under various promoters (UASt, UASp, QUAS, ubiquitin).

Main Results:

• Successfully established a Fly-FUCCI system capable of distinguishing G1, S, and G2 phases in Drosophila.
• Demonstrated the utility of Fly-FUCCI in various tissues, including cultured cells, imaginal discs, salivary glands, and adult midgut.
• Developed a comprehensive toolkit of transgenic lines for diverse research needs.

Conclusions:

• The Fly-FUCCI system provides a valuable method for visualizing cell-cycle dynamics in Drosophila.
• This tool will advance research in Drosophila development, tissue homeostasis, and neoplastic growth.