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Received: 2004-09-22

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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.7 P.673-677


Construction and identification of Fas-targeting siRNA-expressing plasmid

Author(s):  LIU Su-hu, ZHANG Wang-gang, ZHANG Mei, ZHU Qing, TIAN Wei

Affiliation(s):  Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, Department of Clinical Hematology, Second Hospital, Xi’ more

Corresponding email(s):   zhangwanggang2003@yahoo.com

Key Words:  RNA interference, Fas protein, Plasmid, Vector construction

LIU Su-hu, ZHANG Wang-gang, ZHANG Mei, ZHU Qing, TIAN Wei. Construction and identification of Fas-targeting siRNA-expressing plasmid[J]. Journal of Zhejiang University Science B, 2005, 6(7): 673-677.

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author="LIU Su-hu, ZHANG Wang-gang, ZHANG Mei, ZHU Qing, TIAN Wei",
journal="Journal of Zhejiang University Science B",
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%T Construction and identification of Fas-targeting siRNA-expressing plasmid
%A LIU Su-hu
%A ZHANG Wang-gang
%A ZHU Qing
%J Journal of Zhejiang University SCIENCE B
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0673

T1 - Construction and identification of Fas-targeting siRNA-expressing plasmid
A1 - LIU Su-hu
A1 - ZHANG Wang-gang
A1 - ZHANG Mei
A1 - ZHU Qing
A1 - TIAN Wei
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 7
SP - 673
EP - 677
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2005.B0673

Objective: To study the therapeutic potential of Fas inhibition in different diseases, a Fas-targeting siRNA (small interfering)-expressing plasmid was constructed. Methods: The U6 promoter cassette and siFas (small interfering RNA that inhibit Fas expression) template sequence were obtained by PCR method. They were cloned into modified pcDNA3.1. The resultant plasmid pU6-siFas was transfected into P815 cells with lipofectin2000 and selected under G-418-containing culture medium. Fas inhibition in stably transfected cells was detected by immunocytochemistry. Results: The plasmid pU6-siFas efficiently reduced the expression of Fas and conferred G-418 resistance in P815 cells. Conclusion: The successful construction of the siRNA expressing plasmid will facilitate the application of RNA interference technique and lay the foundation for further study of Fas inhibition in the treatment of different diseases such as aplastic anemia and acute liver failure.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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