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Fig. 3 | Cell & Bioscience

Fig. 3

From: A new design of a lentiviral shRNA vector with inducible co-expression of ARGONAUTE 2 for enhancing gene silencing efficiency

Fig. 3

Effect of stem length, loop size, and antisense strand location of shRNA on RNAi efficiency. a Design of a set of shEGFP variants with different stem lengths. The stem lengths were indicated in the name of each shRNA. All the constructs were designed to target EGFP, starting at the position of 417 of EGPF with the stem sequence extending to the 3′ direction. The hairpin loop was the same (5′-TTCAAGAGA-3′) for all of the vectors; b comparison of the activities of shRNAs with different stem lengths in HEK293 cells. G/R fluorescence intensity ratio was measured from the cells co-transfected with pENTR/CMV-EGFP, pDsRed2-c1, shRNA vector with or without AGO2 overexpression vector. shCon/control group was used to normalize the silencing efficiency; c design of 19-bp hairpin stem of shEGFP417 at the orientation of Sense-Loop-Antisense (S-L-AS) or Antisense-Loop-Sense (AS-L-S) structure with three kinds of loop sequences, L1 (4-nt, TTCG), L2, (6-nt, CTCGAG) or L3 (9-nt, TTCAAGAGA); d comparison of the activities of S-L-AS and AS-L-S shRNA with different loop lengths in the presence or absence of AGO2 co-expression. ShCon/control group was used for the normalization of EGFP silencing activities. The results shown are mean ± SD from four independent experiments. ***P < 0.001

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