skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Chirality Dependent Potency Enhancement and Structural Impact of Glycol Nucleic Acid Modification on siRNA

Authors:
ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [2];  [1];  [2];  [1]
  1. Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, United States
  2. Vanderbilt University School of Medicine, Department of Biochemistry, Nashville, Tennessee 37232, United States
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
INDUSTRY
OSTI Identifier:
1374634
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of the American Chemical Society; Journal Volume: 139; Journal Issue: 25
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Schlegel, Mark K., Foster, Donald J., Kel’in, Alexander V., Zlatev, Ivan, Bisbe, Anna, Jayaraman, Muthusamy, Lackey, Jeremy G., Rajeev, Kallanthottathil G., Charissé, Klaus, Harp, Joel, Pallan, Pradeep S., Maier, Martin A., Egli, Martin, and Manoharan, Muthiah. Chirality Dependent Potency Enhancement and Structural Impact of Glycol Nucleic Acid Modification on siRNA. United States: N. p., 2017. Web. doi:10.1021/jacs.7b02694.
Schlegel, Mark K., Foster, Donald J., Kel’in, Alexander V., Zlatev, Ivan, Bisbe, Anna, Jayaraman, Muthusamy, Lackey, Jeremy G., Rajeev, Kallanthottathil G., Charissé, Klaus, Harp, Joel, Pallan, Pradeep S., Maier, Martin A., Egli, Martin, & Manoharan, Muthiah. Chirality Dependent Potency Enhancement and Structural Impact of Glycol Nucleic Acid Modification on siRNA. United States. doi:10.1021/jacs.7b02694.
Schlegel, Mark K., Foster, Donald J., Kel’in, Alexander V., Zlatev, Ivan, Bisbe, Anna, Jayaraman, Muthusamy, Lackey, Jeremy G., Rajeev, Kallanthottathil G., Charissé, Klaus, Harp, Joel, Pallan, Pradeep S., Maier, Martin A., Egli, Martin, and Manoharan, Muthiah. Mon . "Chirality Dependent Potency Enhancement and Structural Impact of Glycol Nucleic Acid Modification on siRNA". United States. doi:10.1021/jacs.7b02694.
@article{osti_1374634,
title = {Chirality Dependent Potency Enhancement and Structural Impact of Glycol Nucleic Acid Modification on siRNA},
author = {Schlegel, Mark K. and Foster, Donald J. and Kel’in, Alexander V. and Zlatev, Ivan and Bisbe, Anna and Jayaraman, Muthusamy and Lackey, Jeremy G. and Rajeev, Kallanthottathil G. and Charissé, Klaus and Harp, Joel and Pallan, Pradeep S. and Maier, Martin A. and Egli, Martin and Manoharan, Muthiah},
abstractNote = {},
doi = {10.1021/jacs.7b02694},
journal = {Journal of the American Chemical Society},
number = 25,
volume = 139,
place = {United States},
year = {Mon Jun 19 00:00:00 EDT 2017},
month = {Mon Jun 19 00:00:00 EDT 2017}
}
  • Motivated by the promises of gene therapy, there is great interest in developing non-viral lipid-based vectors for therapeutic applications due to their low immunogenicity, low toxicity, ease of production, and the potential of transferring large pieces of DNA into cells. In fact, cationic liposome (CL) based vectors are among the prevalent synthetic carriers of nucleic acids (NAs) currently used in gene therapy clinical trials worldwide. These vectors are studied both for gene delivery with CL-DNA complexes and gene silencing with CL-siRNA (short interfering RNA) complexes. However, their transfection efficiencies and silencing efficiencies remain low compared to those of engineered viralmore » vectors. This reflects the currently poor understanding of transfection-related mechanisms at the molecular and self-assembled levels, including a lack of knowledge about interactions between membranes and double stranded NAs and between CL-NA complexes and cellular components. In this review we describe our recent efforts to improve the mechanistic understanding of transfection by CL-NA complexes, which will help to design optimal lipid-based carriers of DNA and siRNA for therapeutic gene delivery and gene silencing.« less
  • The surface structure and DNA hybridization performance of thiolated single-strand DNA (HS-ssDNA) covalently attached to a maleimide-ethylene glycol disulfide (MEG) monolayer on gold have been investigated. Monolayer immobilization chemistry and surface coverage of reactive ssDNA probes were studied by X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry. Orientation of the ssDNA probes was determined by near-edge X-ray absorption fine structure (NEXAFS). Target DNA hybridization on the DNA-MEG probe surfaces was measured by surface plasmon resonance (SPR) to demonstrate the utility of these probe surfaces for detection of DNA targets from both purified target DNA samples and complex biological mixturesmore » such as blood serum. Data from complementary techniques showed that immobilized ssDNA density is strongly dependent on the spotted bulk DNA concentration and buffer ionic strength. Variation of the immobilized ssDNA density had a profound influence on the DNA probe orientation at the surface and subsequent target hybridization efficiency. With increasing surface probe density, NEXAFS polarization dependence results (followed by monitoring the N 1s {yields} {pi}* transition) indicate that the immobilized ssDNA molecules reorient toward a more upright position on the MEG monolayer. SPR assays of DNA targets from buffer and serum showed that DNA hybridization efficiency increased with decreasing surface probe density. However, target detection in serum was better on the 'high-density' probe surface than on the 'high-efficiency' probe surface. The amounts of target detected for both ssDNA surfaces were several orders of magnitude poorer in serum than in purified DNA samples due to nonspecific serum protein adsorption onto the sensing surface.« less
  • Four new polymers, namely [Ni(-tsgluO)(2,4'-bipy){sub 2}(H{sub 2}O){sub 2}]{sub n}.5nH{sub 2}O (1), [Co(-tsgluO)(2,4'-bipy){sub 2}(H{sub 2}O){sub 2}]{sub n}.5nH{sub 2}O (2), [Ni(-tsgluO)(4,4'-bipy)]{sub n}.0.5nH{sub 2}O (3), and [Co(-tsgluO)(4,4'-bipy)]{sub n}.0.5nH{sub 2}O (4), where tsgluO{sup 2-}=(+)-N-p-tolylsulfonyl-L-glutamate dianion, 2,4'-bipy=2,4'-bipyridine, and 4,4'-bipy=4,4'-bipyridine, have been prepared and structurally characterized. Compounds 1 and 2 are isostructural and mononuclear, and crystallize in the acentric monoclinic space group Cc, forming 1D chain structures. Compound 3 is also mononuclear, but crystallizes in the chiral space group P2{sub 1}, forming a homochiral 2D architecture. In contrast to the other complexes, compound 4 crystallizes in the space group P-1 and is composed of binuclear [Co{submore » 2}O{sub 6}N{sub 2}]{sub n}{sup 4-} units, which give rise to a 2D bilayer framework. Moreover, compounds 1, 2, and 4 self-assemble to form 3D supramolecular structures through {pi}-{pi} stacking and hydrogen-bonding interactions, while compound 3 is further hydrogen-bonded to form 3D frameworks. We have demonstrated the influence of the central metal and bipyridine ligands on the framework chirality of the coordination complexes. - Graphical abstract: Four novel polymers based on a chiral ligand were prepared and structurally characterized; it represents the first series of investigations about the effect of central metals and bipyridine ligands on framework chirality.« less
  • The effects of acriflavin, caffeine, and certain basic dyes on RNA, DNA and protein synthesis, survival, and mutationinduction with uv-exposed bacteria are compared. It is established that the agents exert some action on uv-exposed cells which results in increased mutation frequency response and lethality, as well as blockage of RNA, DNA, and protein synthesis. Acriflavine effects on macromolecular synthesis, mutation induction, and lethality are described. (auth)