1.Inhibition of Luciferase expression in mammalian cells by AAV vector plasmid mediated Luciferase shRNA.
Xin MA ; Xiaochun LU ; Jainqiang PENG ; Shuping TAN ; Zhenhua YUAN ; Fang YIN ; Hong WANG ; Xiaobing WU ; Yunde HOU
Chinese Journal of Experimental and Clinical Virology 2002;16(3):253-255
OBJECTIVEConstructing a plasmid containing the shRNA of luciferase to suppress the expression of luciferase in BHK-21 cell.
METHODSA 334 bp human U6 snRNA promoter was amplified from human genomic DNA by PCR and ligated to a 21 bp reverse repeated motif of luciferase target sequence with 9 bp spacer and AAV plasmid pSNAV. The recombinant pSNAV/U6/Luc plasmid cotransfected with pMAMneoLuc or transfected luciferase cell line to detect the effect of luciferase expression separately.
RESULTSpSNAV/U6/luc suppresses the luciferase expression from pMAMneoLuc by 50% and luciferase cell line by 70%.
CONCLUSIONSThe results showed that the short hairpin RNA of luciferase can efficiently suppress its expression in BHK-21.
Cell Line ; Gene Expression ; Genetic Vectors ; Humans ; Luciferases ; biosynthesis ; genetics ; Plasmids ; genetics ; Promoter Regions, Genetic ; RNA ; genetics ; Transfection
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