Phosphoramidate oligonucleotides as potent antisense molecules in cells and in vivo.

Abstract:

:Antisense oligonucleotides are designed to specifically hybridize to a target messenger RNA (mRNA) and interfere with the synthesis of the encoded protein. Uniformly modified oligonucleotides containing N3'-P5' phosphoramidate linkages exhibit (NP) extremely high-affinity binding to single-stranded RNA, do not induce RNase H activity, and are resistant to cellular nucleases. In the present work, we demonstrate that phosphoramidate oligonucleotides are effective at inhibiting gene expression at the mRNA level, by binding to their complementary target present in the 5'-untranslated region. Their mechanism of action was demonstrated by comparative analysis of three expression systems that differ only by the composition of the oligonucleotide target sequence (HIV-1 polypurine tract or PPT sequence) present just upstream from the AUG codon of the firefly luciferase reporter gene: the experiments have been done on isolated cells using oligonucleotide delivery mediated by cationic molecules or streptolysin O (SLO), and in vivo by oligonucleotide electrotransfer to skeletal muscle. In our experimental system phosphoramidate oligonucleotides act as potent and specific antisense agents by steric blocking of translation initiation; they may prove useful to modulate RNA metabolism while maintaining RNA integrity.

journal_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Faria M,Spiller DG,Dubertret C,Nelson JS,White MR,Scherman D,Hélène C,Giovannangeli C

doi

10.1038/83489

subject

Has Abstract

pub_date

2001-01-01 00:00:00

pages

40-4

issue

1

eissn

1087-0156

issn

1546-1696

journal_volume

19

pub_type

杂志文章
  • Enhancement of antitumor immunity by prolonging antigen presentation on dendritic cells.

    abstract::Vaccination with dendritic cells (DCs) pulsed with antigenic peptides derived from various tumor antigens has great, but as yet significantly unrealized, potential in cancer treatment. Here, we describe a strategy for prolonged presentation of an MHC class I-restricted self-peptide on DCs through linkage of it to a ce...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0202-149

    authors: Wang RF,Wang HY

    更新日期:2002-02-01 00:00:00

  • Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown.

    abstract::A central hurdle in developing small interfering RNAs (siRNAs) as therapeutics is the inefficiency of their delivery across the plasma and endosomal membranes to the cytosol, where they interact with the RNA interference machinery. With the aim of improving endosomal release, a poorly understood and inefficient proces...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.3298

    authors: Wittrup A,Ai A,Liu X,Hamar P,Trifonova R,Charisse K,Manoharan M,Kirchhausen T,Lieberman J

    更新日期:2015-08-01 00:00:00

  • Microarray reality checks in the context of a complex disease.

    abstract::A problem in analyzing microarray-based gene expression data is the separation of genes causally involved in a disease from innocent bystander genes, whose expression levels have been secondarily altered by primary changes elsewhere. To investigate this issue systematically in the context of a class of complex human d...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt965

    authors: Miklos GL,Maleszka R

    更新日期:2004-05-01 00:00:00

  • The human use of humanoid beings: chimeras and patent law.

    abstract::As biotechnology advances, the day may soon come for the creation of a self-aware, human-nonhuman chimera. The USPTO has ruled on whether a patent may issue on such an organism, but Congress must still legislate a dividing line between human and non-human patentable subject matter. ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0506-517

    authors: Rabin S

    更新日期:2006-05-01 00:00:00

  • Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells.

    abstract::Previous reports have demonstrated the growth of undifferentiated human embryonic stem (HES) cells on mouse embryonic fibroblast (MEF) feeders and on laminin- or Matrigel-coated plastic surfaces supplemented with MEF-conditioned medium. These xenosupport systems run the risk of cross-transfer of animal pathogens from ...

    journal_title:Nature biotechnology

    pub_type:

    doi:10.1038/nbt726

    authors: Richards M,Fong CY,Chan WK,Wong PC,Bongso A

    更新日期:2002-09-01 00:00:00

  • In vivo delivery of siRNA to immune cells by conjugation to a TLR9 agonist enhances antitumor immune responses.

    abstract::Efficient delivery of small interfering (si)RNA to specific cell populations in vivo remains a formidable challenge to its successful therapeutic application. We show that siRNA synthetically linked to a CpG oligonucleotide agonist of toll-like receptor (TLR)9 targets and silences genes in TLR9(+) myeloid cells and B ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.1564

    authors: Kortylewski M,Swiderski P,Herrmann A,Wang L,Kowolik C,Kujawski M,Lee H,Scuto A,Liu Y,Yang C,Deng J,Soifer HS,Raubitschek A,Forman S,Rossi JJ,Pardoll DM,Jove R,Yu H

    更新日期:2009-10-01 00:00:00

  • Encapsulated cell technology.

    abstract::The potential therapeutic applications of encapsulated cells are enormous. In the US alone, it has been estimated that nearly half-a-trillion dollars are spent each year to care for patients who suffer tissue loss or dysfunction. Over 6 million patients suffer from neurodegenerative disorders such as Alzheimer's disea...

    journal_title:Nature biotechnology

    pub_type: 杂志文章,评审

    doi:10.1038/nbt0996-1107

    authors: Lanza RP,Hayes JL,Chick WL

    更新日期:1996-09-01 00:00:00

  • Lipid production in Nannochloropsis gaditana is doubled by decreasing expression of a single transcriptional regulator.

    abstract::Lipid production in the industrial microalga Nannochloropsis gaditana exceeds that of model algal species and can be maximized by nutrient starvation in batch culture. However, starvation halts growth, thereby decreasing productivity. Efforts to engineer N. gaditana strains that can accumulate biomass and overproduce ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.3865

    authors: Ajjawi I,Verruto J,Aqui M,Soriaga LB,Coppersmith J,Kwok K,Peach L,Orchard E,Kalb R,Xu W,Carlson TJ,Francis K,Konigsfeld K,Bartalis J,Schultz A,Lambert W,Schwartz AS,Brown R,Moellering ER

    更新日期:2017-07-01 00:00:00

  • Hearts and minds.

    abstract::The nonprofit Pew Initiative on Food and Biotechnology is closing, but the need for an independent and neutral body to facilitate dialog on US biotech policy has never been greater. ...

    journal_title:Nature biotechnology

    pub_type: 社论

    doi:10.1038/nbt0207-143

    authors:

    更新日期:2007-02-01 00:00:00

  • Development and function of human innate immune cells in a humanized mouse model.

    abstract::Mice repopulated with human hematopoietic cells are a powerful tool for the study of human hematopoiesis and immune function in vivo. However, existing humanized mouse models cannot support development of human innate immune cells, including myeloid cells and natural killer (NK) cells. Here we describe two mouse strai...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.2858

    authors: Rongvaux A,Willinger T,Martinek J,Strowig T,Gearty SV,Teichmann LL,Saito Y,Marches F,Halene S,Palucka AK,Manz MG,Flavell RA

    更新日期:2014-04-01 00:00:00

  • Chaperones as thermodynamic sensors of drug-target interactions reveal kinase inhibitor specificities in living cells.

    abstract::The interaction between the HSP90 chaperone and its client kinases is sensitive to the conformational status of the kinase, and stabilization of the kinase fold by small molecules strongly decreases chaperone interaction. Here we exploit this observation and assay small-molecule binding to kinases in living cells, usi...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.2620

    authors: Taipale M,Krykbaeva I,Whitesell L,Santagata S,Zhang J,Liu Q,Gray NS,Lindquist S

    更新日期:2013-07-01 00:00:00

  • Metalloenzyme nitrile hydratase: structure, regulation, and application to biotechnology.

    abstract::Nitrile hydratase (NHase), which catalyzes the hydration of nitriles to amides, has been used in the industrial production of acrylamide and nicotinamide. Recent studies on NHases, which are roughly classified into iron and cobalt types according to the metal involved, have clarified the photoactivation mechanism, the...

    journal_title:Nature biotechnology

    pub_type: 杂志文章,评审

    doi:10.1038/nbt0898-733

    authors: Kobayashi M,Shimizu S

    更新日期:1998-08-01 00:00:00

  • Blockade of CD2-LFA-3 interactions protects human skin allografts in immunodeficient mouse/human chimeras.

    abstract::A human skin allograft injury model in immunodeficient mice, engrafted with human peripheral blood mononuclear cells from a different donor, has been used to test whether reagents that block human T cell CD2 interactions with its principal ligand, LFA-3 (CD58), can inhibit immune reactions in vivo. In this model, huma...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0897-759

    authors: Sultan P,Schechner JS,McNiff JM,Hochman PS,Hughes CC,Lorber MI,Askenase PW,Pober JS

    更新日期:1997-08-01 00:00:00

  • The functional landscape of the human phosphoproteome.

    abstract::Protein phosphorylation is a key post-translational modification regulating protein function in almost all cellular processes. Although tens of thousands of phosphorylation sites have been identified in human cells, approaches to determine the functional importance of each phosphosite are lacking. Here, we manually cu...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/s41587-019-0344-3

    authors: Ochoa D,Jarnuczak AF,Viéitez C,Gehre M,Soucheray M,Mateus A,Kleefeldt AA,Hill A,Garcia-Alonso L,Stein F,Krogan NJ,Savitski MM,Swaney DL,Vizcaíno JA,Noh KM,Beltrao P

    更新日期:2020-03-01 00:00:00

  • A functional screen in yeast for regulators and antagonizers of heterologous protein tyrosine kinases.

    abstract::Tyrosine phosphorylation exerts a pivotal role in cell regulation processes of higher eukaryotes. Tight control of the activity of protein tyrosine kinases is crucial for ordered phosphorylation to occur. We have developed a functional screen for tyrosine kinase regulators using c-Src, the first cellular protein tyros...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0596-600

    authors: Superti-Furga G,Jönsson K,Courtneidge SA

    更新日期:1996-05-01 00:00:00

  • Milking the genome for profit.

    abstract::Despite recent accomplishments, significant challenges remain before cloning can become a routine part of transgenic animal production. ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/80231

    authors: Dove A

    更新日期:2000-10-01 00:00:00

  • A catalog of the mouse gut metagenome.

    abstract::We established a catalog of the mouse gut metagenome comprising ∼2.6 million nonredundant genes by sequencing DNA from fecal samples of 184 mice. To secure high microbiome diversity, we used mouse strains of diverse genetic backgrounds, from different providers, kept in different housing laboratories and fed either a ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.3353

    authors: Xiao L,Feng Q,Liang S,Sonne SB,Xia Z,Qiu X,Li X,Long H,Zhang J,Zhang D,Liu C,Fang Z,Chou J,Glanville J,Hao Q,Kotowska D,Colding C,Licht TR,Wu D,Yu J,Sung JJ,Liang Q,Li J,Jia H,Lan Z,Tremaroli V,Dworzyn

    更新日期:2015-10-01 00:00:00

  • Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates.

    abstract::The reactive thiol in cysteine is used for coupling maleimide linkers in the generation of antibody conjugates. To assess the impact of the conjugation site, we engineered cysteines into a therapeutic HER2/neu antibody at three sites differing in solvent accessibility and local charge. The highly solvent-accessible si...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.2108

    authors: Shen BQ,Xu K,Liu L,Raab H,Bhakta S,Kenrick M,Parsons-Reponte KL,Tien J,Yu SF,Mai E,Li D,Tibbitts J,Baudys J,Saad OM,Scales SJ,McDonald PJ,Hass PE,Eigenbrot C,Nguyen T,Solis WA,Fuji RN,Flagella KM,Patel D,Spe

    更新日期:2012-01-22 00:00:00

  • Author Correction: Plants with genetically encoded autoluminescence.

    abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    journal_title:Nature biotechnology

    pub_type: 已发布勘误

    doi:10.1038/s41587-020-0578-0

    authors: Mitiouchkina T,Mishin AS,Somermeyer LG,Markina NM,Chepurnyh TV,Guglya EB,Karataeva TA,Palkina KA,Shakhova ES,Fakhranurova LI,Chekova SV,Tsarkova AS,Golubev YV,Negrebetsky VV,Dolgushin SA,Shalaev PV,Shlykov D,Melnik OA,

    更新日期:2020-08-01 00:00:00

  • An integrated mass spectrometric and computational framework for the analysis of protein interaction networks.

    abstract::Biological systems are controlled by protein complexes that associate into dynamic protein interaction networks. We describe a strategy that analyzes protein complexes through the integration of label-free, quantitative mass spectrometry and computational analysis. By evaluating peptide intensity profiles throughout t...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt1289

    authors: Rinner O,Mueller LN,Hubálek M,Müller M,Gstaiger M,Aebersold R

    更新日期:2007-03-01 00:00:00

  • TagGraph reveals vast protein modification landscapes from large tandem mass spectrometry datasets.

    abstract::Although mass spectrometry is well suited to identifying thousands of potential protein post-translational modifications (PTMs), it has historically been biased towards just a few. To measure the entire set of PTMs across diverse proteomes, software must overcome the dual challenges of covering enormous search spaces ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/s41587-019-0067-5

    authors: Devabhaktuni A,Lin S,Zhang L,Swaminathan K,Gonzalez CG,Olsson N,Pearlman SM,Rawson K,Elias JE

    更新日期:2019-04-01 00:00:00

  • Programmable RNA editing by recruiting endogenous ADAR using engineered RNAs.

    abstract::Current tools for targeted RNA editing rely on the delivery of exogenous proteins or chemically modified guide RNAs, which may lead to aberrant effector activity, delivery barrier or immunogenicity. Here, we present an approach, called leveraging endogenous ADAR for programmable editing of RNA (LEAPER), that employs s...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/s41587-019-0178-z

    authors: Qu L,Yi Z,Zhu S,Wang C,Cao Z,Zhou Z,Yuan P,Yu Y,Tian F,Liu Z,Bao Y,Zhao Y,Wei W

    更新日期:2019-09-01 00:00:00

  • A large peptidome dataset improves HLA class I epitope prediction across most of the human population.

    abstract::Prediction of HLA epitopes is important for the development of cancer immunotherapies and vaccines. However, current prediction algorithms have limited predictive power, in part because they were not trained on high-quality epitope datasets covering a broad range of HLA alleles. To enable prediction of endogenous HLA ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/s41587-019-0322-9

    authors: Sarkizova S,Klaeger S,Le PM,Li LW,Oliveira G,Keshishian H,Hartigan CR,Zhang W,Braun DA,Ligon KL,Bachireddy P,Zervantonakis IK,Rosenbluth JM,Ouspenskaia T,Law T,Justesen S,Stevens J,Lane WJ,Eisenhaure T,Lan Zhang G,

    更新日期:2020-02-01 00:00:00

  • A recombinant polypeptide extends the in vivo half-life of peptides and proteins in a tunable manner.

    abstract::Increasing the in vivo residence times of protein therapeutics could decrease their dosing frequencies. We show that genetic fusion of an unstructured recombinant polypeptide of 864 amino acids, called XTEN, to a peptide or protein provides an apparently generic approach to extend plasma half-life. Allometric scaling ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.1588

    authors: Schellenberger V,Wang CW,Geething NC,Spink BJ,Campbell A,To W,Scholle MD,Yin Y,Yao Y,Bogin O,Cleland JL,Silverman J,Stemmer WP

    更新日期:2009-12-01 00:00:00

  • A policy approach to the development of molecular diagnostic tests.

    abstract::Efficiently generating evidence of clinical utility is a major challenge for ensuring clinical adoption of valuable diagnostics. A new approach to reimbursement in the United States offers a balance between evidence and incentives for molecular diagnostic tests. ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt1110-1157

    authors: Schulman KA,Tunis SR

    更新日期:2010-11-01 00:00:00

  • Isolation of neuronal precursors by sorting embryonic forebrain transfected with GFP regulated by the T alpha 1 tubulin promoter.

    abstract::Neuronal precursor cells are widespread in the forebrain ventricular/subventricular zone, and may provide a cellular substrate for brain repair. Clonal lines derived from single progenitors can become progressively less representative of their parental precursors with time and passage in vitro. We have developed an al...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0298-196

    authors: Wang S,Wu H,Jiang J,Delohery TM,Isdell F,Goldman SA

    更新日期:1998-02-01 00:00:00

  • Publisher Correction: Single-cell RNA-seq analysis software providers scramble to offer solutions.

    abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章,已发布勘误

    doi:10.1038/s41587-020-0510-7

    authors: Eisenstein M

    更新日期:2020-05-01 00:00:00

  • Prospects for plant-derived antibacterials.

    abstract::Can weakly active phytochemicals be combined synergistically to produce new antibacterial treatments? ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt1206-1504

    authors: Lewis K,Ausubel FM

    更新日期:2006-12-01 00:00:00

  • Targeted RNA sequencing reveals the deep complexity of the human transcriptome.

    abstract::Transcriptomic analyses have revealed an unexpected complexity to the human transcriptome, whose breadth and depth exceeds current RNA sequencing capability. Using tiling arrays to target and sequence select portions of the transcriptome, we identify and characterize unannotated transcripts whose rare or transient exp...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt.2024

    authors: Mercer TR,Gerhardt DJ,Dinger ME,Crawford J,Trapnell C,Jeddeloh JA,Mattick JS,Rinn JL

    更新日期:2011-11-13 00:00:00

  • Synthetic CD4 exocyclics inhibit binding of human immunodeficiency virus type 1 envelope to CD4 and virus replication in T lymphocytes.

    abstract::CD4 functions as a major T-cell surface receptor for human immunodeficiency virus by binding the human immunodeficiency virus type 1 (HIV-1) envelope protein gp120 with relatively high affinity. We have developed constrained aromatically modified analogs of the secondary structures of the first domain of CD4 in order ...

    journal_title:Nature biotechnology

    pub_type: 杂志文章

    doi:10.1038/nbt0297-150

    authors: Zhang X,Gaubin M,Briant L,Srikantan V,Murali R,Saragovi U,Weiner D,Devaux C,Autiero M,Piatier-Tonneau D,Greene MI

    更新日期:1997-02-01 00:00:00