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In vivo GFP interference

The in vivo GFP interference (iGFPi) method can be used to knock down expression of proteins tagged with EGFP and other GFP variants using RNA interference constructs.

iGFPi illustration originating in Jose Pastor-Pareja and Tian Xu (2011), Shaping Cells and Organs in Drosophila by Opposing Roles of Fat Body-Secreted Collagen IV and Perlecan. Dev. Cell 21(2): 245--256

P{UAS-GFP.dsRNA.R} effectively knocks down expression of EGFP, but has not been tested with other GFP variants. It will not knock down expression of YFP or other non-GFP fluorescent proteins. Its use was described by Roignant et al. (2003) and Pastor-Pareja and Xu (2011).

The VALIUM-based "EGFP.shRNA" constructs were described by Neumuller (2012). They are effective against EGFP and EGFP derivatives (like EYFP) but are not effective against normal GFP or GFP derivatives. More information on these insertions, including the primer sequences used to make the constructs and estimates of the relative levels of knockdown can be found here in csv form or as an Excel file.

Lines for RNAi of other fluors can be found on the RNAi for non-fly genes page.
Go to the RNAi home page.
Go to the UAS home page.

  Gene(s) Stock # Vector Insertion Chr. Genotype
1 EGFP 9331 P{UAS-GFP.dsRNA.R}143 2 w[1118]; P{w[+mC]=UAS-GFP.dsRNA.R}143
2 EGFP 44412 P{UAS-GFP.dsRNA.R}143 2 w[*]; P{w[+mC]=UAS-GFP.dsRNA.R}143/CyO; Sb[1]/TM6B, Tb[1]
3 EGFP 44414 P{UAS-GFP.dsRNA.R}143 2 w[*]; P{w[+mC]=UAS-GFP.dsRNA.R}143; P{w[+mC]=He-GAL4.Z}85, P{w[+mC]=UAS-myr-mRFP}2/TM6B, Tb[1]
4 EGFP 9330 P{UAS-GFP.dsRNA.R}142 3 w[1118]; P{w[+mC]=UAS-GFP.dsRNA.R}142
5 EGFP 44410 P{UAS-GFP.dsRNA.R}142 3 w[*]; P{w[+m*]=GAL4-ey.H}4-8/CyO; P{w[+mC]=UAS-GFP.dsRNA.R}142
6 EGFP 44415 P{UAS-GFP.dsRNA.R}142 3 w[*]; wg[Sp-1]/CyO; P{w[+mC]=UAS-GFP.dsRNA.R}142
7 EGFP 63147 P{UAS-GFP.dsRNA.R}142 3 w[*]; P{w[+mC]=Cg-GAL4.A}2, P{w[+mC]=UAS-myr-mRFP}1/CyO; P{w[+mC]=UAS-GFP.dsRNA.R}142/TM6B, Tb[1]
8 EGFP 63148 P{UAS-GFP.dsRNA.R}142 3 w[*]; P{w[nub.PK]=nub-GAL4.K}2, P{w[+mC]=UAS-myr-mRFP}1/CyO; P{w[+mC]=UAS-GFP.dsRNA.R}142/TM6B, Tb[1]
9 EGFP, EYFP 41552 VALIUM20 P{VALIUM20-EGFP.shRNA.4}attP40 2 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.4}attP40
10 EGFP, EYFP 41555 VALIUM20 P{VALIUM20-EGFP.shRNA.1}attP40 2 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.1}attP40
11 EGFP, EYFP 41559 VALIUM20 P{VALIUM20-EGFP.shRNA.3}attP40 2 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.3}attP40
12 EGFP, EYFP 41553 VALIUM20 P{VALIUM20-EGFP.shRNA.4}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.4}attP2
13 EGFP, EYFP 41554 VALIUM20 P{VALIUM20-EGFP.shRNA.2}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.2}attP2
14 EGFP, EYFP 41556 VALIUM20 P{VALIUM20-EGFP.shRNA.1}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.1}attP2
15 EGFP, EYFP 41560 VALIUM20 P{VALIUM20-EGFP.shRNA.3}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM20-EGFP.shRNA.3}attP2
16 EGFP, EYFP 66387 VALIUM20 P{VALIUM20-EGFP.shRNA.1}attP2 3 w[*]; P{w[+mC]=Ubi-GFP(S65T)nls}2L, P{w[+mC]=UAS-RFP.W}2; P{y[+t7.7 v[+t1.8]=VALIUM20-EGFP.shRNA.1}attP2
17 EGFP, EYFP 41550 VALIUM22 P{VALIUM22-EGFP.shRNA.4}attP40 2 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM22-EGFP.shRNA.4}attP40
18 EGFP, EYFP 41557 VALIUM22 P{VALIUM22-EGFP.shRNA.1}attP40 2 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM22-EGFP.shRNA.1}attP40
19 EGFP, EYFP 41551 VALIUM22 P{VALIUM22-EGFP.shRNA.4}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM22-EGFP.shRNA.4}attP2
20 EGFP, EYFP 41558 VALIUM22 P{VALIUM22-EGFP.shRNA.1}attP2 3 y[1] sc[*] v[1]; P{y[+t7.7] v[+t1.8]=VALIUM22-EGFP.shRNA.1}attP2