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Figure 2 | BMC Biotechnology

Figure 2

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

Figure 2

Structure and knockdown efficacy of CMV-shLuc with different stem length and polyadenylation (pA) or U1 transcription termination signals (U1). (a) Predicted stem-loop structure of CMV-shLuc with different stem lengths (20, 21, 25 and 29 bp). Guide strand is highlighted in grey (b) Luciferase knockdown by CMV-shLuc20, CMV-shLuc21, CMV-shLuc25, CMV-shLuc29 with pA transcription termination signal. (c) Luciferase knockdown by CMV-shLuc20, CMV-shLuc21, CMV-shLuc25, CMV-shLuc29 with U1 transcription termination signal. 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 short hairpin expressing plasmid. Firefly luciferase expression was normalized to Renilla luciferase expression. H1-shLuc was used as a positive control. H1-shScr and CMV-shScr 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 < 0.05, **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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