[The simultaneous knock-down of Ku70 and Ku80 by a tandem Ku-shRNA-encoding plasmid expression system].
OBJECTIVE
To extend the use of vector-derived siRNA by generating multiple shRNAs in the same plasmid.


METHODS
Construct a vector that expresses shRNAs targeting on Ku70 and Ku80 in tandem. The gene silencing efficiency of each shRNA was verified previously. After identification by restriction digestion and DNA sequencing, the reconstructed plasmid, named psiRNAKus, was transfected into the human hepatoma cell line HepG2. The tandem-shRNA-induced silencing of targeted genes was determined by RT-PCR at RNA level and Western blot at protein level.


RESULTS
The shRNAs encoded by psiRNAKus down-regulated both the expression of Ku70 and Ku80.


CONCLUSIONS
The vector-derived siRNA delivery system that allows multiple shRNA species to be expressed from the same vector may be of value in experimental and therapeutic applications.
Ren
Jing-hua
JH
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Lin
Ju-sheng
JS
Chang
Ying
Y
Wu
Ying
Y
Zhang
Ying-hui
YH
Zhang
Qiang
Q
He
Xing-xing
XX
D034961:Antigens, Nuclear / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D055785:Gene Knockdown Techniques; D056945:Hep G2 Cells; D006801:Humans; D000072200:Ku Autoantigen; D010957:Plasmids; D012313:RNA / Q000235:genetics*; D034741:RNA, Small Interfering
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D034741:RNA, Small Interfering; D012313:RNA; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen