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- W2387470434 abstract "Objective To construct a retrovirus-mediated expression system containing short hairpin RNA(shRNA) targeting mouse RANK gene,and to study its influence on the osteoclastogenesis of mouse bone marrow-derived macrophages.Methods A recombinant retrovirus vector pSUPER-shRANK was generated by cloning shRNA targeting mouse RANK gene into pSuper-Retro vector.To obtain retrovirus supernatant,pSUPER-retro-shRANK and packaging plasmid vector(PIK) were transfected into 293 FT packaging cells by calcium phosphate transfection method.BMMs were repeatedly infected by the retrovirus supernatant,and stable BMM clones were generated after screening with puromycin.RANK proteins were then analyzed by Western blot.BMMs and infected BMMs were induced by 100 ng/ml RANKL and 30 ng/ml M-CSF for nine days.Osteoclasts and bone resorption pits were detected by TRAP staining and Scanning Electron Microscopy.Results The pSUPER-shRANK recombinant retrovirus vector had been constructed correctly by restriction enzyme digestion and sequencing methods assays.Stable infected BMMs clones were generated after screening with puromycin.The expression of RANK protein in infected BMMs reduced significantly(80.7%),compared with uninfected BMMs(P0.01).Nine days later,there was a reduction in the number of TRAP+ multinucleated cells in infected BMMs(6.80±1.64) compared with uninfected BMMs(82.00±4.64)(P0.05).Furthermore there was significant reduction in resorption areas on bone slices in infected BMMs(2.80%±0.84%) compared with uninfected BMMs(16.60%±2.70%)(P0.05),indicating profound inhibition of osteoclastic bone resorption.Conclusions The pSUPER-shRANK retrovirus vector showed effective inhibition on the expression of RANK at protein levels;the osteoclastogenesis of BMMs was inhibited by RNAi targeting RANK." @default.
- W2387470434 created "2016-06-24" @default.
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- W2387470434 date "2011-01-01" @default.
- W2387470434 modified "2023-09-25" @default.
- W2387470434 title "The effect of retrovirus-mediated shRNA targeting mouse RANK on osteoclastogenesis of bone marrow-derived macrophages" @default.
- W2387470434 hasPublicationYear "2011" @default.
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