RNA interference by small hairpin RNAs synthesised under control of the human 7S K RNA promoter

被引:26
作者
Koper-Emde, D [1 ]
Herrmann, L [1 ]
Sandrock, B [1 ]
Benecke, BJ [1 ]
机构
[1] Ruhr Univ Bochum, Fak Chem, Lehrstuhl Biochem 1, D-44780 Bochum, Germany
关键词
gene silencing; lamin A/C; pol III transcription; siRNA;
D O I
10.1515/BC.2004.103
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Small interfering RNAs (siRNAs) represent RNA duplexes of 21 nucleotides in length that inhibit gene expression. We have used the human geneexternal 7S K RNA promoter for synthesis of short hairpin RNAs (shRNAs) which efficiently target human lamin mRNA via RNA interference (RNAi). Here we demonstrate that orientation of the target sequence within the shRNA construct is important for interference. Furthermore, effective interference also depends on the length and/or structure of the shRNA. Evidence is presented that the human 7S K promoter is more active in vivo than other geneexternal promoters, such as the human U6 small nuclear RNA (snRNA) gene promoter.
引用
收藏
页码:791 / 794
页数:4
相关论文
共 18 条
[1]   NUCLEOTIDE-SEQUENCES IN XENOPUS 5S DNA REQUIRED FOR TRANSCRIPTION TERMINATION [J].
BOGENHAGEN, DF ;
BROWN, DD .
CELL, 1981, 24 (01) :261-270
[2]   FUNCTIONAL REDUNDANCY OF PROMOTER ELEMENTS ENSURES EFFICIENT TRANSCRIPTION OF THE HUMAN 7SK GENE IN-VIVO [J].
BOYD, DC ;
TURNER, PC ;
WATKINS, NJ ;
GERSTER, T ;
MURPHY, S .
JOURNAL OF MOLECULAR BIOLOGY, 1995, 253 (05) :677-690
[3]   Efficient transcription of the EBER2 gene depends on the structural integrity of the RNA [J].
Dümpelmann, E ;
Mittendorf, H ;
Benecke, BJ .
RNA, 2003, 9 (04) :432-442
[4]   Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells [J].
Elbashir, SM ;
Harborth, J ;
Lendeckel, W ;
Yalcin, A ;
Weber, K ;
Tuschl, T .
NATURE, 2001, 411 (6836) :494-498
[5]  
Harborth J, 2001, J CELL SCI, V114, P4557
[6]   TRANSCRIPTION OF HUMAN 7S K DNA INVITRO AND INVIVO IS EXCLUSIVELY CONTROLLED BY AN UPSTREAM PROMOTER [J].
KLEINERT, H ;
BENECKE, BJ .
NUCLEIC ACIDS RESEARCH, 1988, 16 (04) :1319-1331
[7]   EXPRESSION OF A HUMAN 7S-K RNA GENE INVIVO REQUIRES A NOVEL POL-III UPSTREAM ELEMENT [J].
KLEINERT, H ;
BREDOW, S ;
BENECKE, BJ .
EMBO JOURNAL, 1990, 9 (03) :711-718
[8]   TRANSCRIPTION OF A HUMAN U6 SMALL NUCLEAR-RNA GENE INVIVO WITHSTANDS DELETION OF INTRAGENIC SEQUENCES BUT NOT OF AN UPSTREAM TATATA BOX [J].
KUNKEL, GR ;
PEDERSON, T .
NUCLEIC ACIDS RESEARCH, 1989, 17 (18) :7371-7379
[9]  
KUNKEL GR, 1992, J BIOL CHEM, V267, P14250
[10]   Effective expression of small interfering RNA in human cells [J].
Paul, CP ;
Good, PD ;
Winer, I ;
Engelke, DR .
NATURE BIOTECHNOLOGY, 2002, 20 (05) :505-508