Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (7): 1267-1270.doi: 10.3969/j.issn.1673-8225.2012.07.030

Previous Articles     Next Articles

Construction and identification of a lentiviral vector for RNA interference of suppressors of cytokine signaling 1  

Zhou Min, Qian Yan-rong, Zhang Qiu-rui, Cheng Ting, Wan Huan-ying   

  1. Department of Respiratory Medicine,  Ruijin Hospital, Shanghai Jiao Tong University, Shanghai  200025, China
  • Received:2011-09-07 Revised:2011-11-11 Online:2012-02-12 Published:2012-02-12
  • Contact: Wan Huan-ying, Master, Chief physician, Department of Respiratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai 200025, China hy_wan2006@yahoo.com.cn
  • About author:Zhou Min☆, Doctor, Associate chief physician, Department of Respiratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai 200025, China doctor_zhou_99@yahoo.com.cn
  • Supported by:

    Key Project of Pharmaceutical Science and Technology Agency of Shanghai, No. 09411951400*

PDF

374

Abstract:

BACKGROUND: The previous studies mainly used liposome transfection or other vectors for research of suppressors of cytokine signaling 1 (SOCS1). However, those transfections have the disadvantages of low efficiency and less security.
OBJECTIVE: To construct a lentiviral vector for RNA interference of SOCS1 gene and identify it. 
METHODS: Three pairs of complementary short hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene were designed, and then were inserted into pPll3.7 vectors by gene recombination technology. The recombinant plasmids with correct sequence were cotransfected with packaging plasmids into 293T cells. The cell culture supernatant was obtained after 48 hours in order to transfect A549 cells. The expression of SOCS1 in target cells was detected by western-blot.
RESULTS AND CONCLUSION: The shRNA had been confirmed to insert into pPll3.7 vectors by sequencing of the recombinant plasmids. The western-blot detection showed that the expression of SOCS1 in A549 cells was inhibited after the cells were successfully transfected by the lentiviral vector supernatant. A lentiviral vector for RNA interference of SOCS1 gene was successfully constructed in this study.

CLC Number: