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公开(公告)号：EP1407044A2
公开(公告)日：2004-04-14
申请号：EP01985833.1
申请日：2001-11-29
申请人： Max-Planck-Gesellschaft zur Förderungder Wissenschaften e.V. , Europäisches Laboratorium für Molekularbiologie
IPC分类号： C12Q1/68
CPC分类号： C12N15/113 , A01K2217/075 , A61K9/0019 , A61K38/00 , A61K48/00 , C12N15/1079 , C12N15/111 , C12N2310/14 , C12N2310/321 , C12N2310/53 , C12N2330/30 , C12N2310/3521
摘要： Double-stranded RNA (dsRNA) induces sequence-specific post-transcriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 19-23 nt short RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the produced siRNP complex.
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2.发明公开
公开(公告)号：EP1520022A2
公开(公告)日：2005-04-06
申请号：EP03763812.9
申请日：2003-07-10
发明人： TUSCHL, Thomas , MARTINEZ, Javier , PATKANIOWSKA, Agnieszka , URLAUB, Henning , LÜHRMANN, Reinhard
CPC分类号： C12N15/113 , A61K47/549 , A61K47/557 , C07K14/4705 , C07K19/00 , C12N15/111 , C12N2310/14 , C12N2310/351 , C12N2310/3513 , C12N2320/30 , C12N2320/51 , C12N2330/30
摘要： The present invention relates to sequence and structural features of single-stranded (ss) RNA molecules required to mediate target-specific nucleic acid modifications by RNA-interference (RNAi), such as target mRNA degradation and/or DNA methylation.
摘要翻译： 本发明涉及通过RNA干扰（RNAi）介导靶特异性核酸修饰所需的单链（ss）RNA分子的序列和结构特征，例如靶mRNA降解和/或DNA甲基化。
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公开(公告)号：EP1309726A2
公开(公告)日：2003-05-14
申请号：EP01922870.9
申请日：2001-03-30
申请人： WHITEHEAD INSTITUTE FOR BIOMEDICAL RESEARCH , Max-Planck-Gesellschaft zur Förderungder Wissenschaften e.V. , Massachusetts Institute of Technology , UNIVERSITY OF MASSACHUSETTS MEDICAL CENTER
IPC分类号： C12Q1/68
CPC分类号： C12N15/113 , A01K67/0336 , A01K2207/05 , A01K2217/075 , A01K2227/703 , A01K2267/03 , A61K38/00 , C07H21/02 , C12N15/1079 , C12N15/111 , C12N2310/14 , C12N2310/321 , C12N2310/53 , C12N2330/30 , C12Q1/66 , C12N2310/3521
摘要： The present invention relates to a Drosophila in vitro system which was used to demonstrate that dsRNA is processed to RNA segments 21-23 nucleotides (nt) in length. Furthermore, when these 21-23 nt fragments are purified and added back to Drosophila extracts, they mediate RNA interference in the absence of long dsRNA. Thus, these 21-23 nt fragments are the sequence-specific mediators of RNA degradation. A molecular signal, which may be their specific length, must be present in these 21-23 nt fragments to recruit cellular factors involved in RNAi. This present invention encompasses these 21-23 nt fragments and their use for specifically inactivating gene function. The use of these fragments (or chemically synthesized oligonucleotides of the same or similar nature) enables the targeting of specific mRNAs for degradation in mammalian cells, where the use of long dsRNAs to elicit RNAi is usually not practical, presumably because of the deleterious effects of the interferon response. This specific targeting of a particular gene function is useful in functional genomic and therapeutic applications.
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公开(公告)号：EP3199631B1
公开(公告)日：2019-01-30
申请号：EP17160119.8
申请日：2001-11-29
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公开(公告)号：EP1407044B1
公开(公告)日：2007-09-19
申请号：EP01985833.1
申请日：2001-11-29
申请人： Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. , Europäisches Laboratorium für Molekularbiologie
IPC分类号： C12Q1/68
CPC分类号： C12N15/113 , A01K2217/075 , A61K9/0019 , A61K38/00 , A61K48/00 , C12N15/1079 , C12N15/111 , C12N2310/14 , C12N2310/321 , C12N2310/53 , C12N2330/30 , C12N2310/3521
摘要： Double-stranded RNA (dsRNA) induces sequence-specific post-transcriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 19-23 nt short RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the produced siRNP complex.
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公开(公告)号：EP1407044B2
公开(公告)日：2017-11-15
申请号：EP01985833.1
申请日：2001-11-29
申请人： Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. , Europäisches Laboratorium für Molekularbiologie
IPC分类号： C12N15/113
CPC分类号： C12N15/113 , A01K2217/075 , A61K9/0019 , A61K38/00 , A61K48/00 , C12N15/1079 , C12N15/111 , C12N2310/14 , C12N2310/321 , C12N2310/53 , C12N2330/30 , C12N2310/3521
摘要： Double-stranded RNA (dsRNA) induces sequence-specific post-transcriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 19-23 nt short RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the produced siRNP complex.
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公开(公告)号：EP3199631A1
公开(公告)日：2017-08-02
申请号：EP17160119.8
申请日：2001-11-29
申请人： Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. , Europäisches Laboratorium für Molekularbiologie (EMBL)
CPC分类号： C12N15/113 , A01K2217/075 , A61K9/0019 , A61K38/00 , A61K47/549 , A61K48/00 , C12N15/1079 , C12N15/111 , C12N2310/14 , C12N2310/321 , C12N2310/53 , C12N2330/30 , C12N2310/3521
摘要： Double-stranded RNA (dsRNA) induces sequence-specific post-transcriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 19-23 nt short RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the produced siRNP complex.
摘要翻译： 双链RNA（dsRNA）通过称为RNA干扰（RNAi）的过程在许多生物体中诱导序列特异性转录后基因沉默。 使用果蝇体外系统，我们证明19-23 nt短的RNA片段是RNAi的序列特异性介质。 短干扰RNA（siRNA）由来自长dsRNA的RNase III样加工反应产生。 具有突出3'末端的化学合成的siRNA双链体在裂解物中介导有效靶RNA裂解，并且裂解位点位于导向siRNA跨越的区域的中心附近。 此外，我们提供的证据表明，dsRNA加工的方向决定了正义或反义目标RNA是否可以被产生的siRNP复合物切割。
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