Stocks for isolating RNA from specific cells

Translating Ribosomal Affinity Purification (TRAP)

Thomas et al. (2012) created stocks for expressing a GFP-tagged ribosomal protein under UAS control. Ribosomes from GAL4-expressing cells can be recovered by immunoprecipitation with an anti-GFP antibody and actively translated mRNAs isolated.

Stock #Genotype
42681w[*]; P{UAS-GFP-RpL10Ab}BF2b
42682P{UAS-GFP-RpL10Ab}BF24, w[*]; l(2)*[*]/CyO
42683w[*]; P{UAS-GFP-RpL10Ab}BF27

TU-tagging and EC-tagging

Miller et al. (2009) and Hida et al., 2017 generated stocks that express either uracil phosphoribosyltransferase (UPRT) or a cytosine deaminase (CD) – UPRT fusion under UAS control. In flies fed 4-thiouracil (4TU), 4TU is modified by UPRT and incorporated into newly synthesized RNA (TU-tagging). In flies fed 5-ethynylcytosine (5EC), 5EC is modified by CD:UPRT and incorporated into newly synthesized RNA (EC-tagging) . Thio- or ethynyl-labeled RNA can be selectively recovered by affinity purification.

Stock #Genotype
77119y[1] w[*]; P{UAS-HA-CD:UPRT.n}attP40
77120w[*]; wg[Sp-1]/CyO, P{ActGFP}JMR1; P{UAS-HA-CD:UPRT.n}attP2
77121y[1] w[*]; P{UAS-HA-CD:UPRT.o}attP40
77122y[1] w[*]; P{UAS-HA-CD:UPRT.o}attP2
27602w[*]; P{UAS-HA-UPRT}2.1; Dr[1]/TM3, Sb[1]
27603w[*]; wg[Sp-1]/CyO; P{UAS-UPRT.M}3.1
27604w[*]; wg[Sp-1]/CyO; P{UAS-HA-UPRT}3.2