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Figure 1 | BMC Biotechnology

Figure 1

From: Optimization and comparison of knockdown efficacy between polymerase II expressed shRNA and artificial miRNA targeting luciferase and Apolipoprotein B100

Figure 1

Structure and knockdown efficacy of short hairpins targeting luciferase (shLuc) with different TSS and different TTS. (a) Schematic representation of shLuc constructs expressed from H1 and CMV promoters. Different TTS (pA, T5 and U1) are presented. Hairpin location (+5 or +6) is shown with reference to the TSS (set as +1). The shLuc consists of 20 nt perfectly complementary hairpin structure and a loop of 9 nt. (b) Luciferase knockdown by CMV-shLuc with different TSS and different TTS. Renilla and Firefly luciferase were measured two days post-transfection with 2,5 ng Firefly luciferase reporter, 0,5 ng Renilla luciferase and 100 ng shLuc expressing plasmids. Firefly luciferase expression was normalized to Renilla luciferase expression. H1-shLuc was used as a positive control, H1-shGFP and CMV-shGFP served as negative controls and were set at 100%. Data are represented as mean values ± SD from three independent experiments analyzed with the factor correction method [19]. **p < 0.01 versus negative control One-way ANOVA test with Bonferroni post test.

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BMC Biotechnology

ISSN: 1472-6750

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