Uas yfp rab5

Manufactured by Bloomington Drosophila Stock Center

The UAS-YFP-Rab5 is a genetically encoded fluorescent protein that can be used to visualize the localization and dynamics of the small GTPase Rab5, which is involved in early endosome formation and trafficking. This tool allows researchers to study the subcellular distribution and trafficking of Rab5 in live cells.

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4 protocols using uas yfp rab5

Drosophila Strains for Cell Organelle Tracking

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Flies were crossed, reared and kept on standard food (Meidi LLC) at 25°C. The following Drosophila strains used in this study had been previously generated by us (Z.H.’s lab): Hand-GFP (expressing green fluorescent protein in the nuclei of nephrocytes and cardiomyocytes; Han and Olson, 2005 (link); Huang et al., 2023 (link)), and Mhc-ANF-RFP (expressing ANF red fluorescent protein in muscle myosin heavy chain promoter-driven atrial natriuretic peptide-red fluorescent protein in muscle cells; Zhang et al., 2013 (link)). The following lines were obtained from the Bloomington Drosophila Stock Center (BDSC; Indiana University, IN): Dot-Gal4 (ID 6903; Ugt36A1 driver), tub-Gal4 (ID 30029; alphaTub84B driver), UAS-YFP-Rab5 (ID 24616), UAS-YFP-Rab7 (ID 23270), UAS-YFP-Rab11 (ID 9790), and UAS-mCherry-Atg8a (ID 37750). As control, w¹¹¹⁸ (BDSC; ID 3605) flies were used in the crosses.

Zhu J.Y., Lee J.G., Fu Y., van de Leemput J., Ray P.E, & Han Z. (2023). APOL1-G2 accelerates nephrocyte cell death by inhibiting the autophagy pathway. Disease Models & Mechanisms, 16(12), dmm050223.

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Drosophila Neurodegenerative Pathway Imaging

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Flies were crossed, reared, and kept on standard food (Meidi LLC) at 22°C under standard conditions. The following Drosophila lines were obtained from the Bloomington Drosophila Stock Center (BDSC; Indiana University, IN): Dot-Gal4 (ID 6903; Ugt36A1 driver), UAS-YFP-Rab5 (ID 24616), UAS-YFP-Rab7 (ID 23270), UAS-mCherry-Atg8a (ID 37750), and w¹¹¹⁸ (BDSC; ID 3605) which were used in the crosses. The Hand-GFP line (express green fluorescent protein in the nuclei of nephrocytes and cardiomyocytes) was previously generated by our team ^{31 (link)}.

Zhu J.Y., Fu Y., van de Leemput J., Yu Y., Li J., Ray P.E, & Han Z. (2024). HIV-1 Nef acts in synergy with APOL1-G1 to induce nephrocyte cell death in a new Drosophila model of HIV-related kidney diseases. bioRxiv.

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Drosophila Genetic Toolkit for Rab5 Study

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Rab5^Z1 was an EMS allele generated in our lab; Rab5² was from Marcos González; UAS-Rab5 GFP/TM3, UAS-Rab5 RNAi/CyO, and UAS-Rab5 RNAi/TM3 were from Xinhua Lin; C587-GAL4 was from Ting Xie; nosGAL4VP16 was from Ruth Lehmann; puc^E69 (puc-lacZ) was from Joaquim Culi; kek1-lacZ is an enhancer trap line (BB142) from T. Schüpbach; UAS-YFP-Rab5 (BL24616), UAS-TSG101-RNAi (BL35710), UAS-Vps25-RNAi (BL26286), UAS-avl-RNAi (BL29546), UAS-Cdc42-RNAi (BL28021), dpp^P10638 (dpp-lacZ), tj-GAL4, tub-GAL80^ts, UAS-dpp-RNAi (BL25782), UAS-bsk^DN, UAS-hep^CA, and 2L deficiency flies were from Bloomington Drosophila Stock Center.
Fly stocks were maintained under standard culture conditions and all flies were dissected 0–2 days after eclosion unless otherwise indicated. For GAL4/GAL80^ts controlled RNAi or gene expression, flies were raised at 18 or 25°, shifted to 29° upon eclosion, and aged at 29° for 5 days before dissection. Germline or somatic cyst cell specific clones were generated by expressing FLP with nosGAL4VP16 or C587GAL4 (Kawase et al. 2004 (link)), respectively.

Tang Y., Geng Q., Chen D., Zhao S., Liu X, & Wang Z. (2017). Germline Proliferation Is Regulated by Somatic Endocytic Genes via JNK and BMP Signaling in Drosophila. Genetics, 206(1), 189-197.

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Drosophila Endocytic Trafficking Regulators

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All crosses and staging were performed at 25°C. Stocks are described in FlyBase (http://flybase.bio.indiana.edu). UAS-YFP-Rab11, UAS-YFP-Rab11^Q70L, UAS-YFP-Rab11^S25N, UAS-Rab11-RNAi, UAS-Cdc42^V12, UAS-YFP-Rab8^Q67L, UAS-YFP-Rab4, UAS-YFP-Rab14^Q97L, UAS-aPKC-RNAi, UAS-Rho1, Dia⁵ FRT40A, UAS-YFP-Rab5, GMR-gal4, Nuf^EY11342, Rho1^72F, Cdc42⁴ FRT19A, UAS-Sec8, Rab11^EP3017, UAS-Rok-CAT, UAS-LIMK-HA, UAS-MBS-RNAi, and Sqh^AX3; RLC-GFP were provided by the Bloomington Stock Center (Indiana University, Bloomington, IN); UAS-Rab5-RNAi was provided by the Vienna Drosophila RNAi Center (Vienna, Austria); UAS-Rok-RNAi and UAS-PKN-RNAi were provided by the National Institute of Genetics (Mishima, Japan); Shg^R69 FRT42D was provided by C. Micchelli (Washington University, St. Louis, MO); Par6^Δ226 FRT19A and UAS-aPKC^CAAX were provided by C. Doe (University of Oregon, Eugene, OR); Sec5^E10 FRT40A and UAS-Sec5 were provided by T. L. Schwarz (Harvard Medical School, Boston, MA); dRip11^d00190 was provided by the Exelixis Collection (Harvard Medical School); and UAS-Rho1 RNAi, UAS-Cdc42 RNAi, and UAS-Cdc42 were generated by our lab (Warner and Longmore, 2009a (link), b (link)).

Yashiro H., Loza A.J., Skeath J.B, & Longmore G.D. (2014). Rho1 regulates adherens junction remodeling by promoting recycling endosome formation through activation of myosin II. Molecular Biology of the Cell, 25(19), 2956-2969.

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