MCFO - MultiColor FlpOut
Updated November 12, 2015
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MCFO (MultiColor FlpOut) is a system for multicolor stochastic labeling of cells. Multiple color combinations are achieved by the use of five different tags (HA, V5, OLLAS, FLAG and Myc) each in a backbone of non-fluorescent GFP (in FlyBase symbols this is referred to as smGdp for 'Green darkened Protein'). Spatial and temporal control of expression is regulated thru the use of one of two binary expression systems (GAL4/UAS or lexA/lexAop) and excision of an intervening stop cassette by one of three recombinases (FLP, B3R or KDR).



An example of staining in the lamina and medulla from Nern et al.
MCFO example

For more information see: Nern et al., 2015. PNAS. 112(22):E2967.

Stocks carrying a single insertion.

Stk # Promoter Cassette Tag(s) Chr Genotype
62104 alphaTub84B B3RT GAL80 3 w[1118]; PBac{y[+mDint2] w[+mC]=tubP-(B3RT.GAL80)}VK00027
62103 alphaTub84B FRT GAL80 3 w[1118]; PBac{y[+mDint2] w[+mC]=tubP-(FRT.GAL80)}VK00027
62105 alphaTub84B KDRT GAL80 3 w[1118]; PBac{y[+mDint2] w[+mC]=tubP-(KDRT.GAL80)}VK00027
62106 lexAop FRT HA 3 w[1118]; PBac{y[+mDint2] w[+mC]=13xLexAop2(FRT.stop)myr::smGdP-HA}VK00005
62108 lexAop FRT OLLAS 3 w[1118]; P{y[+t7.7] w[+mC]=13xLexAop2(FRT.stop)myr::smGdP-OLLAS}attP2
62107 lexAop FRT V5 2 w[1118]; P{y[+t7.7] w[+mC]=13xLexAop2(FRT.stop)myr::smGdP-V5}attP40
62111 lexAop KDRT FLAG 2 w[1118]; P{y[+t7.7] w[+mC]=13xLexAop2(KDRT.stop)myr::smGdP-FLAG}su(Hw)attP5
62109 lexAop KDRT HA 3 w[1118]; PBac{y[+mDint2] w[+mC]=13xLexAop2(KDRT.stop)myr::smGdP-HA}VK00005
62112 lexAop KDRT OLLAS 3 w[1118]; P{y[+t7.7] w[+mC]=13xLexAop2(KDRT.stop)myr::smGdP-OLLAS}attP2
62110 lexAop KDRT V5 3 w[1118]; P{y[+t7.7] w[+mC]=13xLexAop2(KDRT.stop)myr::smGdP-V5}su(Hw)attP1
62132 UAS B3RT HA 3 w[1118]; PBac{y[+mDint2] w[+mC]=10xUAS(B3RT.stop)myr::smGdP-HA}VK00005
62135 UAS B3RT OLLAS 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(B3RT.stop)myr::smGdP-OLLAS}attP2
62137 UAS B3RT OLLAS 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(KDRT.stop)myr::smGdP-OLLAS}attP2
62134 UAS B3RT V5 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(B3RT.stop)myr::smGdP-V5}su(Hw)attP5
62133 UAS B3RT V5 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(B3RT.stop)myr::smGdP-V5}su(Hw)attP1
62136 UAS B3RT V5 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(KDRT.stop)myr::smGdP-V5}su(Hw)attP1
62130 UAS FRT FLAG 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-FLAG}attP40
62122 UAS FRT FLAG & Myc 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-FLAG-THS-10xUAS(FRT.stop)myr::smGdP-cMyc}su(Hw)attP5
62121 UAS FRT FLAG & Myc 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-FLAG-THS-10xUAS(FRT.stop)myr::smGdP-cMyc}su(Hw)attP1
62128 UAS FRT GFP & HA 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS-IVS-myr::sfGFP-THS-10xUAS(FRT.stop)myr::smGdP-HA}attP40
62127 UAS FRT GFP & HA 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS-IVS-myr::sfGFP-THS-10xUAS(FRT.stop)myr::smGdP-HA}attP2
62126 UAS FRT HA 3 w[1118]; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005
62125 UAS FRT Myc 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-cMyc}attP40
62131 UAS FRT OLLAS 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-OLLAS}attP2
62129 UAS FRT V5 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5}attP40
62124 UAS FRT V5 & HA 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP5
62123 UAS FRT V5 & HA 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
62120 UAS FRT V5 & Myc 2 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-cMyc}su(Hw)attP5
62119 UAS FRT V5 & Myc 3 w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-cMyc}su(Hw)attP1

Stocks carrying combinations of insertions

All cassettes in these stocks are FRT-based (there are no B3RT or KDRT cassettes). Most also express a FLP variant.

Stk # FLP construct Tags a.k.a. Genotype
64092 w[1118] P{y[+t7.7] w[+mC]=10xUAS-IVS-myr::smGdP-HA}attP18 P{y[+t7.7] w[+mC]=13xLexAop2-IVS-myr::smGdP-V5}su(Hw)attP8; TM2/TM6B, Tb[1]
64093 HA, V5, FLAG w[1118]; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64094 HA, V5, FLAG, OLLAS w[1118]; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-OLLAS}attP2 PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64085 hs-FLPG5 HA, V5, FLAG MCFO-1 w[1118] P{y[+t7.7] w[+mC]=hs-FLPG5.PEST}attP3; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64086 hs-FLPG5 HA, V5, FLAG, OLLAS MCFO-2 w[1118] P{y[+t7.7] w[+mC]=hs-FLPG5.PEST}attP3; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-OLLAS}attP2 PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64087 57C10-FLPL HA, V5, FLAG MCFO-3 w[1118] P{y[+t7.7] w[+mC]=GMR57C10-FLPL}su(Hw)attP8; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64088 57C10-FLPG5 HA, V5, FLAG MCFO-4 w[1118] P{y[+t7.7] w[+mC]=GMR57C10-FLPG5}su(Hw)attP8; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64089 57C10-FLPG5.PEST HA, V5, FLAG MCFO-5 w[1118] P{y[+t7.7] w[+mC]=GMR57C10-FLPG5.PEST}su(Hw)attP8; PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64090 57C10-FLPL HA, V5, FLAG, OLLAS MCFO-6 w[1118] P{y[+t7.7] w[+mC]=GMR57C10-FLPL}su(Hw)attP8; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-OLLAS}attP2 PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1
64091 57C10-FLPG5.PEST HA, V5, FLAG, OLLAS MCFO-7 w[1118] P{y[+t7.7] w[+mC]=GMR57C10-FLPG5.PEST}attP18; P{y[+t7.7] w[+mC]=10xUAS(FRT.stop)myr::smGdP-OLLAS}attP2 PBac{y[+mDint2] w[+mC]=10xUAS(FRT.stop)myr::smGdP-HA}VK00005 P{10xUAS(FRT.stop)myr::smGdP-V5-THS-10xUAS(FRT.stop)myr::smGdP-FLAG}su(Hw)attP1