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Fly-FUCCI: Fluorescent Ubiquitination-based Cell Cycle Indicator

Cell cycle phases can be identified using combinations of two fusion proteins.

Figure of cell cycle marker expression from Zielke et al. (2014), Fly-FUCCI - a versatile tool for studying cell proliferation in complex tissues. Cell Reports.

In the example above, GFP is fused to the portion of the E2f protein targeted by CLR4Cdt2 (also known as l(2)dtl) for degradation in S phase, and mRFP is fused to the portion of the CycB protein targeted by the Anaphase Promoting Complex/cyclosome for degradation during G1.

In the stocks listed below, UAST, UASp, QUAS or ubiquitin regulatory sequences drive expression of E2f or CycB degrons fused to fluorescent proteins. Some of the fusion proteins carry nuclear localization sequences, but the non-NLS fusion proteins tend to work better in endoreplicating cells.

Figure of cell cycle marker expression from Zielke et al. (2014), Fly-FUCCI - a versatile tool for studying cell proliferation in complex tissues. Cell Reports.

This method is described in Zielke et al. (2014), Fly-FUCCI: a versatile tool for studying cell proliferation in complex tissues.

FUCCI# Promoter E2F tag CycB tag Chr Stk# Genotype
#1 Ubi GFP mRFP 2 55098 w[1118]; P{w[+mC]=Ubi-GFP.E2f1.1-230}19 P{w[+mC]=Ubi-mRFP1.CycB.1-266}12/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#2 Ubi GFP mRFP 3 55099 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=Ubi-GFP.E2f1.1-230}5 P{w[+mC]=Ubi-mRFP1.CycB.1-266}4/TM6B, Tb[1]
#3 UASp GFP mRFP 2 55100 w[1118]; P{w[+mC]=UASp-GFP.E2f1.1-230}26 P{w[+mC]=UASp-mRFP1.CycB.1-266}81; MKRS/TM6B, Tb[+]
#4 UASp GFP mRFP 3 55101 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UASp-GFP.E2f1.1-230}64 P{w[+mC]=UASp-mRFP1.CycB.1-266}21/TM6B, Tb[1]
#5 UAS CFP Venus 2 55102 w[1118]; P{w[+mC]=UAS-CFP.E2f1.1-230}4 P{w[+mC]=UAS-Venus.CycB.1-266}7; MKRS/TM6B, Tb[1]
#6 UAS CFP Venus 3 55103 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UAS-CFP.E2f1.1-230}3 P{w[+mC]=UAS-Venus.CycB.1-266}1
#7 UASp CFP Venus 2 55104 w[1118]; P{w[+mC]=UASp-CFP.E2f1.1-230}4 P{w[+mC]=UASp-Venus.CycB.1-266}10/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#8 UASp CFP Venus 3 55105 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UASp-CFP.E2f1.1-230}1 P{w[+mC]=UASp-Venus.CycB.1-266}1/TM6B, Tb[1]
#9 QUAS CFP Venus 2 55106 w[1118]; P{w[+mC]=QUAS-CFP.E2f1.1-230}5 P{w[+mC]=QUAS-Venus.CycB.1-266}21/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#10 QUAS CFP Venus 3 55107 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=QUAS-CFP.E2f1.1-230}1 P{w[+mC]=QUAS-Venus.CycB.1-266}10
#11 QUAS GFP mRFP 2 55108 w[1118]; P{w[+mC]=QUAS-GFP.E2f1.1-230}2 P{w[+mC]=QUAS-mRFP1.CycB.1-266}4; MKRS/TM6B, Tb[1]
#12 QUAS GFP mRFP 3 55109 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=QUAS-GFP.E2f1.1-230}1 P{w[+mC]=QUAS-mRFP1.CycB.1-266}3/TM6B, Tb[1]
#13 UASp GFP mRFP.NLS 2 55110 w[1118]; P{w[+mC]=UASp-GFP.E2f1.1-230}26 P{w[+mC]=UASp-mRFP1.NLS.CycB.1-266}4/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#14 UASp GFP mRFP.NLS 3 55111 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UASp-GFP.E2f1.1-230}64 P{w[+mC]=UASp-mRFP1.NLS.CycB.1-266}5/TM6B, Tb[1]
#15 UAS CFP Venus.NLS 2 55112 w[1118]; P{w[+mC]=UAS-CFP.E2f1.1-230}6 P{w[+mC]=UAS-Venus.NLS.CycB.1-266}38/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#16 UAS CFP Venus.NLS 3 55303 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UAS-CFP.E2f1.1-230}3 P{w[+mC]=UAS-Venus.NLS.CycB.1-266}3/TM6B, Tb[1]
#17 UASp CFP Venus.NLS 2 55113 w[1118]; P{w[+mC]=UASp-CFP.E2f1.1-230}4 P{w[+mC]=UASp-Venus.NLS.CycB.1-266}1/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#18 UASp CFP Venus.NLS 3 55114 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UASp-CFP.E2f1.1-230}1 P{w[+mC]=UASp-Venus.NLS.CycB.1-266}5/TM6B, Tb[1]
#19 QUAS CFP Venus.NLS 2 55115 w[1118]; P{w[+mC]=QUAS-CFP.E2f1.1-230}5 P{w[+mC]=QUAS-Venus.NLS.CycB.1-266}7/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#20 QUAS CFP Venus.NLS 3 55116 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=QUAS-CFP.E2f1.1-230}1 P{w[+mC]=QUAS-Venus.NLS.CycB.1-266}30/TM6B, Tb[1]
#21 UAS GFP mRFP 2 55117 w[1118]; P{w[+mC]=UAS-GFP.E2f1.1-230}32 P{w[+mC]=UAS-mRFP1.CycB.1-266}13/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#22 UAS GFP mRFP 3 55118 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UAS-GFP.E2f1.1-230}26 P{w[+mC]=UAS-mRFP1.CycB.1-266}5/TM6B, Tb[1]
#23 QUAS GFP mRFP.NLS 2 55119 w[1118]; P{w[+mC]=QUAS-GFP.E2f1.1-230}2 P{w[+mC]=QUAS-mRFP1.NLS.CycB.1-266}3B; MKRS/TM6B, Tb[1]
#24 QUAS GFP mRFP.NLS 3 55120 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=QUAS-GFP.E2f1.1-230}1 P{w[+mC]=QUAS-mRFP1.NLS.CycB.1-266}8/TM6B, Tb[1]
#25 UAS GFP mRFP.NLS 2 55121 w[1118]; P{w[+mC]=UAS-GFP.E2f1.1-230}32 P{w[+mC]=UAS-mRFP1.NLS.CycB.1-266}19/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#26 UAS GFP mRFP.NLS 3 55122 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=UAS-GFP.E2f1.1-230}26 P{w[+mC]=UAS-mRFP1.NLS.CycB.1-266}17/TM6B, Tb[1]
#27 Ubi GFP mRFP.NLS 2 55123 w[1118]; P{w[+mC]=Ubi-GFP.E2f1.1-230}19 P{w[+mC]=Ubi-mRFP1.NLS.CycB.1-266}15/CyO, P{ry[+t7.2]=en1}wg[en11]; MKRS/TM6B, Tb[1]
#28 Ubi GFP mRFP.NLS 3 55124 w[1118]; Kr[If-1]/CyO, P{ry[+t7.2]=en1}wg[en11]; P{w[+mC]=Ubi-GFP.E2f1.1-230}5 P{w[+mC]=Ubi-mRFP1.NLS.CycB.1-266}12/TM6B, Tb[1]