%0 Journal Article
%T Inhibiting GDF-8 Expression by Retrovirus-Based RNAi Stably<br>逆转录病毒载体介导的RNA干扰稳定抑制肌肉生长抑制素GDF-8的表达
%A Chaowu Liu
%A Zhuo Yang
%A Bin Zhao
%A Changmei Liu
%A <br>刘超武
%A 杨 倬
%A 赵 斌
%A 刘长梅
%J 生物工程学报
%D 2008
%I 
%X We cloned human U6 promoter from pAVU6 + 27 vector into pXSN to transcripe small RNA. Meanwhile, a shRNA targeting GDF-8 was cloned down-stream of the hU6 promoter to construct recombinant vector. Then the packing cell GP-293 was co-transfected the recombinant with pVSV-G to gernarate virus particle. Resistant C2C12 cell pools were screened using G418. Levels of mRNA and protein of GDF-8 were tested by Real-Time PCR and western blotting. Cell proliferation and cell cycle were analyzed using MTT and FACS. The expression of GDF-8 was dramatically decreased by the retrovirus-based system in C2C12 cells. Cells proliferated effectively after integrating the recombinant. The cells in G0/G1 phase decreased by 13.7%, while cells in S phase increased by 14.9%. In conclusion, the retrovirus-based RNAi could be used to stably silence GDF-8. It can be a powerful tool in curing muscle atrophy.
%K GDF-8 gene
%K C2C12 cells
%K retrovirus vector
%K shRNA<br>肌肉生长抑制素GDF-8
%K C2C12细胞系
%K 逆转录病毒
%K shRNA
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=B3B2278CD48A7460AF93F1F85DC12ABC&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=0B39A22176CE99FB&sid=1D01216AD76577EC&eid=627456E7977439A4&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=18