Author: Liu, Hong Yan; Gao, Xiaohu
Title: A Universal Protein Tag for Delivery of SiRNA-Aptamer Chimeras Document date: 2013_11_7
ID: 0atfsivf_24
Snippet: Functional characterization of siRNA-aptamer chimera. To test the functionality of the siRNA block, the chimera described above and GFP siRNA control (Qiagen) at a final concentration of 50 nM were transfected into C4-2 prostate cancer cells stably expressing GFP using Lipofectamine RNAi MAX (Invitrogen) following the instructions provided by the manufacturer. To evaluate the targeting specificity of the aptamer block, PSMA-positive LNCaP cells a.....
Document: Functional characterization of siRNA-aptamer chimera. To test the functionality of the siRNA block, the chimera described above and GFP siRNA control (Qiagen) at a final concentration of 50 nM were transfected into C4-2 prostate cancer cells stably expressing GFP using Lipofectamine RNAi MAX (Invitrogen) following the instructions provided by the manufacturer. To evaluate the targeting specificity of the aptamer block, PSMA-positive LNCaP cells and PSMA-negative PC3 cells were treated with complex of chimera and dsRBD-His 18 (chimera/protein tag molar ratio at 152, 100 nM chimera) in serum free medium for 2 hours, followed by incubation in complete medium for another 12 h. DAPI (30 nM) was added to stain cell nuclei. Fluorescent images were captured on an Olympus IX-71 inverted microscope equipped with 5 long-pass filters and a colored CCD camera.
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