A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity

被引：285

作者：

Bramsen, Jesper B. ^{[1
]}

Laursen, Maria B. ^{[1
]}

Nielsen, Anne F. ^{[1
]}

Hansen, Thomas B. ^{[1
]}

Bus, Claus ^{[1
]}

Langkjaer, Niels ^{[2
]}

Babu, B. Ravindra ^{[2
]}

Hojland, Torben ^{[2
]}

Abramov, Mikhail ^{[3
]}

Van Aerschot, Arthur ^{[3
]}

Odadzic, Dalibor ^{[4
]}

Smicius, Romualdas ^{[4
]}

Haas, Jens ^{[4
]}

Andree, Cordula ^{[5
]}

Barman, Jharna ^{[6
]}

Wenska, Malgorzata ^{[6
]}

Srivastava, Puneet ^{[6
]}

Zhou, Chuanzheng ^{[6
]}

Honcharenko, Dmytro ^{[6
]}

Hess, Simone ^{[7
]}

Mueller, Elke ^{[7
]}

Bobkov, Georgii V. ^{[8
]}

Mikhailov, Sergey N. ^{[8
]}

Fava, Eugenio ^{[5
]}

Meyer, Thomas F. ^{[7
]}

Chattopadhyaya, Jyoti ^{[6
]}

Zerial, Marino ^{[5
]}

Engels, Joachim W. ^{[4
]}

Herdewijn, Piet ^{[3
]}

Wengel, Jesper ^{[2
]}

Kjems, Jorgen ^{[1
]}

机构：

[1] Univ Aarhus, Dept Mol Biol, DK-8000 Aarhus C, Denmark

[2] Univ So Denmark, Nucle Acid Ctr, Odense, Denmark

[3] Katholieke Univ Leuven, Rega Inst Med Res, Louvain, Belgium

[4] Goethe Univ Frankfurt, Inst Organ Chem & Chem Biol, Frankfurt, Germany

[5] Max Planck Inst Mol Cell Biol & Genet, Dresden, Germany

[6] Uppsala Univ, Dept Bioorgan Chem, Biomed Ctr, Uppsala, Sweden

[7] Max Planck Inst Infect Biol, Dept Mol Biol, Berlin, Germany

[8] Russian Acad Sci, Engelhardt Inst Mol Biol, Moscow, Russia

来源：

NUCLEIC ACIDS RESEARCH | 2009年 / 37卷 / 09期

基金：

新加坡国家研究基金会;

关键词：

LOCKED NUCLEIC-ACID; SHORT INTERFERING RNA; PASSENGER-STRAND; STRUCTURAL BASIS; BUILDING-BLOCKS; GUIDE-STRAND; IN-VITRO; ARGONAUTE2; OLIGONUCLEOTIDES; NUCLEOSIDES;

D O I：

10.1093/nar/gkp106

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：

The use of chemically synthesized short interfering RNAs (siRNAs) is currently the method of choice to manipulate gene expression in mammalian cell culture, yet improvements of siRNA design is expectably required for successful application in vivo. Several studies have aimed at improving siRNA performance through the introduction of chemical modifications but a direct comparison of these results is difficult. We have directly compared the effect of 21 types of chemical modifications on siRNA activity and toxicity in a total of 2160 siRNA duplexes. We demonstrate that siRNA activity is primarily enhanced by favouring the incorporation of the intended antisense strand during RNA-induced silencing complex (RISC) loading by modulation of siRNA thermodynamic asymmetry and engineering of siRNA 3-overhangs. Collectively, our results provide unique insights into the tolerance for chemical modifications and provide a simple guide to successful chemical modification of siRNAs with improved activity, stability and low toxicity.

引用

页码：2867 / 2881

页数：15

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