siRNA carrying an (E)-vinylphosphonate moiety at the 5′ end of the guide strand augments gene silencing by enhanced binding to human Argonaute-2

被引:65
作者
Elkayam, Elad [1 ,2 ,3 ]
Parmar, Rubina [4 ]
Brown, Christopher R. [4 ]
Willoughby, Jennifer L. [4 ]
Theile, Christopher S. [4 ]
Manoharan, Muthiah [4 ]
Joshua-Tor, Leemor [1 ,2 ,3 ]
机构
[1] Keck Struct Biol Lab, Cold Spring Harbor, NY 11724 USA
[2] Howard Hughes Med Inst, Cold Spring Harbor, NY 11724 USA
[3] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA
[4] Alnylam Pharmaceut, 300 Third St, Cambridge, MA 02142 USA
关键词
CRYSTAL-STRUCTURE; STRUCTURAL-ANALYSIS; RNAI; INTERFERENCE; EXPRESSION; COMPLEXES; IMPROVE; SYSTEM; DUPLEX; STEPS;
D O I
10.1093/nar/gkw1171
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
Efficient gene silencing by RNA interference (RNAi) in vivo requires the recognition and binding of the 5'-phosphate of the guide strand of an siRNA by the Argonaute protein. However, for exogenous siRNAs it is limited by the rapid removal of the 5'-phosphate of the guide strand by metabolic enzymes. Here, we have determined the crystal structure of human Argonaute-2 in complex with the metabolically stable 5'-(E)-vinylphosphonate (5'-E-VP) guide RNA at 2.5-angstrom resolution. The structure demonstrates how the 5'binding site in the Mid domain of human Argonaute-2 is able to adjust the key residues in the 5'-nucleotide binding pocket to compensate for the change introduced by the modified nucleotide. This observation also explains improved binding affinity of the 5'-E-VP - modified siRNA to human Argonaute-2 in-vitro, as well as the enhanced silencing in the context of the trivalent N-acetylgalactosamine (GalNAc)conjugated siRNA in mice relative to the un-modified siRNA.
引用
收藏
页码:3528 / 3536
页数:9
相关论文
共 48 条
[1]   PHENIX: a comprehensive Python']Python-based system for macromolecular structure solution [J].
Adams, Paul D. ;
Afonine, Pavel V. ;
Bunkoczi, Gabor ;
Chen, Vincent B. ;
Davis, Ian W. ;
Echols, Nathaniel ;
Headd, Jeffrey J. ;
Hung, Li-Wei ;
Kapral, Gary J. ;
Grosse-Kunstleve, Ralf W. ;
McCoy, Airlie J. ;
Moriarty, Nigel W. ;
Oeffner, Robert ;
Read, Randy J. ;
Richardson, David C. ;
Richardson, Jane S. ;
Terwilliger, Thomas C. ;
Zwart, Peter H. .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 :213-221
[2]   Fully 2′-modified oligonucleotide duplexes with improved in vitro potency and stability compared to unmodified small interfering RNA [J].
Allerson, CR ;
Sioufi, N ;
Jarres, R ;
Prakash, TP ;
Naik, N ;
Berdeja, A ;
Wanders, L ;
Griffey, RH ;
Swayze, EE ;
Bhat, B .
JOURNAL OF MEDICINAL CHEMISTRY, 2005, 48 (04) :901-904
[3]   Crystal structure of the MID-PIWI lobe of a eukaryotic Argonaute protein [J].
Boland, Andreas ;
Huntzinger, Eric ;
Schmidt, Steffen ;
Izaurralde, Elisa ;
Weichenrieder, Oliver .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (26) :10466-10471
[4]   Crystal structure and ligand binding of the MID domain of a eukaryotic Argonaute protein [J].
Boland, Andreas ;
Tritschler, Felix ;
Heimstaedt, Susanne ;
Izaurralde, Elisa ;
Weichenrieder, Oliver .
EMBO REPORTS, 2010, 11 (07) :522-527
[5]   RNAi therapeutics: a potential new class of pharmaceutical drugs [J].
Bumcrot, David ;
Manoharan, Muthiah ;
Koteliansky, Victor ;
Sah, Dinah W. Y. .
NATURE CHEMICAL BIOLOGY, 2006, 2 (12) :711-719
[6]   Preclinical Development of a Subcutaneous ALAS1 RNAi Therapeutic for Treatment of Hepatic Porphyrias Using Circulating RNA Quantification [J].
Chan, Amy ;
Liebow, Abigail ;
Yasuda, Makiko ;
Gan, Lin ;
Racie, Tim ;
Maier, Martin ;
Kuchimanchi, Satya ;
Foster, Don ;
Milstein, Stuart ;
Charisse, Klaus ;
Sehgal, Alfica ;
Manoharan, Muthiah ;
Meyers, Rachel ;
Fitzgerald, Kevin ;
Simon, Amy ;
Desnick, Robert J. ;
Querbes, William .
MOLECULAR THERAPY-NUCLEIC ACIDS, 2015, 4
[7]   Real-time quantification of microRNAs by stem-loop RT-PCR [J].
Chen, CF ;
Ridzon, DA ;
Broomer, AJ ;
Zhou, ZH ;
Lee, DH ;
Nguyen, JT ;
Barbisin, M ;
Xu, NL ;
Mahuvakar, VR ;
Andersen, MR ;
Lao, KQ ;
Livak, KJ ;
Guegler, KJ .
NUCLEIC ACIDS RESEARCH, 2005, 33 (20) :e179.1-e179.9
[8]  
Elkayam E, 2012, CELL, V150, P100, DOI 10.1016/j.cell.2012.05.017
[9]   Features and development of Coot [J].
Emsley, P. ;
Lohkamp, B. ;
Scott, W. G. ;
Cowtan, K. .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 2010, 66 :486-501
[10]   The Making of a Slicer: Activation of Human Argonaute-1 [J].
Faehnle, Christopher R. ;
Elkayam, Elad ;
Haase, Astrid D. ;
Hannon, Gregory J. ;
Joshua-Tor, Leemor .
CELL REPORTS, 2013, 3 (06) :1901-1909