![Figure 1. Visualization of gene expression. (A) Schematic representation of the gene expression plasmid (modified from Janicki et al. [2004]). The cassette consists of 256 copies of lac operator repeats, 96 copies of TREs, a minimal CMV promoter, CFP fused to a peroxisomal targeting signal SKL (CFP-SKL), 24 MS2 translational operators (MS2 repeats), a rabbit β-globin intron/exon module, and a cleavage/polyadenylation signal. LacI-mCherry binds to the lac operator repeats resulting in visualization of the DNA. Tet-On (rtTA) expression in the presence of Dox induces gene expression from the minimal CMV promoter. MS2-YFP (YFP fused to the MS2 coat protein) binds to the MS2 repeats, allowing visualization of the transcribed mRNA. (B) 200 copies of the gene expression cassette in A are stably integrated as a transgene array at the 1p36 region in human U2OS cells, referred to as U2OS-2-6-3 cells (Janicki et al., 2004). Transient expression of LacI-mCherry, Tet-On in the presence of Dox, and MS2-YFP in U2OS-2-6-3 cells allowed visualization of DNA (a, arrow), RNA (b, arrow), and CFP-SKL protein (c, blue). Bar, 5 μm.](https://cdn.rupress.org/rup/content_public/journal/jcb/180/1/10.1083_jcb.200706060/8/51fig1.jpeg?Expires=1773501755&Signature=AGSKE3QXw5IkKg8sC2iHTVbhgenbfQutmhdPgoCli1n5tuEf-5WuHrkSUslxxFJgNaAYI3jM4XGM7xDvZVxFE~LHOClbvrJHPE~OagA8qXTmopPairDafRk3rq0bRscEYoH~u0VY1yv3X~BrAkTmsicWYazs-TmFgT2RuXoXOj3Lk5HK93TIGb5ly8QBDckOsZNvEl~1tL~sMdbsvztyBRdEt8m8ybtv4TeG~cdk2u6SHG79IS8~vsrxqGmNCl5wJt5EHnMO97qn~HLOpOgcO98U5ASOK~PaAFK1T0~gntQyPS1tdI6OIdjjTJZY5uaBTs4cJXfVWHxnBsA~7WhZEg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Visualization of gene expression. (A) Schematic representation of the gene expression plasmid (modified from Janicki et al. [2004]). The cassette consists of 256 copies of lac operator repeats, 96 copies of TREs, a minimal CMV promoter, CFP fused to a peroxisomal targeting signal SKL (CFP-SKL), 24 MS2 translational operators (MS2 repeats), a rabbit β-globin intron/exon module, and a cleavage/polyadenylation signal. LacI-mCherry binds to the lac operator repeats resulting in visualization of the DNA. Tet-On (rtTA) expression in the presence of Dox induces gene expression from the minimal CMV promoter. MS2-YFP (YFP fused to the MS2 coat protein) binds to the MS2 repeats, allowing visualization of the transcribed mRNA. (B) 200 copies of the gene expression cassette in A are stably integrated as a transgene array at the 1p36 region in human U2OS cells, referred to as U2OS-2-6-3 cells (Janicki et al., 2004). Transient expression of LacI-mCherry, Tet-On in the presence of Dox, and MS2-YFP in U2OS-2-6-3 cells allowed visualization of DNA (a, arrow), RNA (b, arrow), and CFP-SKL protein (c, blue). Bar, 5 μm.
