Automated design of hammerhead ribozymes and validation by targeting the PABPN1 gene transcript.

Abstract:

:We present a new publicly accessible web-service, RiboSoft, which implements a comprehensive hammerhead ribozyme design procedure. It accepts as input a target sequence (and some design parameters) then generates a set of ranked hammerhead ribozymes, which target the input sequence. This paper describes the implemented procedure, which takes into consideration multiple objectives leading to a multi-objective ranking of the computer-generated ribozymes. Many ribozymes were assayed and validated, including four ribozymes targeting the transcript of a disease-causing gene (a mutant version of PABPN1). These four ribozymes were successfully tested in vitro and in vivo, for their ability to cleave the targeted transcript. The wet-lab positive results of the test are presented here demonstrating the real-world potential of both hammerhead ribozymes and RiboSoft. RiboSoft is freely available at the website http://ribosoft.fungalgenomics.ca/ribosoft/.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kharma N,Varin L,Abu-Baker A,Ouellet J,Najeh S,Ehdaeivand MR,Belmonte G,Ambri A,Rouleau G,Perreault J

doi

10.1093/nar/gkv1111

subject

Has Abstract

pub_date

2016-02-29 00:00:00

pages

e39

issue

4

eissn

0305-1048

issn

1362-4962

pii

gkv1111

journal_volume

44

pub_type

杂志文章
  • Contacts between the growing peptide chain and the 23S RNA in the 50S ribosomal subunit.

    abstract::Peptides of defined length carrying a diazirine photoaffinity label attached either to the alpha-NH2 group of the N-terminal methionine residue, or to the epsilon-NH2 group of an immediately adjacent lysine residue, were prepared in situ on Escherichia coli ribosomes in the presence of a synthetic mRNA analogue. Pepti...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.8.1394

    authors: Stade K,Riens S,Bochkariov D,Brimacombe R

    更新日期:1994-04-25 00:00:00

  • Structure and function of the Toscana virus cap-snatching endonuclease.

    abstract::Toscana virus (TOSV) is an arthropod-borne human pathogen responsible for seasonal outbreaks of fever and meningoencephalitis in the Mediterranean basin. TOSV is a segmented negative-strand RNA virus (sNSV) that belongs to the genus phlebovirus (family Phenuiviridae, order Bunyavirales), encompassing other important h...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz838

    authors: Jones R,Lessoued S,Meier K,Devignot S,Barata-García S,Mate M,Bragagnolo G,Weber F,Rosenthal M,Reguera J

    更新日期:2019-11-18 00:00:00

  • The binding of poly(rA) and poly(rU) to denatured DNA. I. Model studies with homopolymers.

    abstract::We have compared the properties of the poly(rA).oligo(dT) complex with those of the poly(rU).oligo(dA)n complex. Three main differences were found. First, poly(rA) and oligo(dT)n do not form a complex in concentrations of CsCl exceeding 2 M because the poly(rA) is insoluble in high salt. If the complex is made in low ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/3.4.1013

    authors: Mol JN,Borst P

    更新日期:1976-04-01 00:00:00

  • The bacterial and mitochondrial ribosomal A-site molecular switches possess different conformational substates.

    abstract::The A site of the small ribosomal subunit participates in the fidelity of decoding by switching between two states, a resting 'off' state and an active decoding 'on' state. Eight crystal structures of RNA duplexes containing two minimal decoding A sites of the Homo sapiens mitochondrial wild-type, the A1555G mutant or...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkn112

    authors: Kondo J,Westhof E

    更新日期:2008-05-01 00:00:00

  • Single base extension (SBE) with proofreading polymerases and phosphorothioate primers: improved fidelity in single-substrate assays.

    abstract::Model single base extension (SBE) genotyping reactions with individual deoxy-, dideoxy- and acyclonucleoside triphosphates are monitored by MALDI-TOF mass spectrometry. Three non-proofreading DNA polymerases display remarkably high misincorporation (up to 64% of correct incorporation) when extending primers with singl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gng007

    authors: Di Giusto D,King GC

    更新日期:2003-02-01 00:00:00

  • Helicobacter pylori oriC--the first bipartite origin of chromosome replication in Gram-negative bacteria.

    abstract::Binding of the DnaA protein to oriC leads to DNA melting within the DNA unwinding element (DUE) and initiates replication of the bacterial chromosome. Helicobacter pylori oriC was previously identified as a region localized upstream of dnaA and containing a cluster of DnaA boxes bound by DnaA protein with a high affin...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks742

    authors: Donczew R,Weigel C,Lurz R,Zakrzewska-Czerwinska J,Zawilak-Pawlik A

    更新日期:2012-10-01 00:00:00

  • Real-time measurement of in vitro transcription.

    abstract::We have developed a simple method to measure RNA synthesis in real time. In this technique, transcription reactions are performed in the presence of molecular beacons that possess a 2'-O-methylribonucleotide backbone. These probes become fluorescent as they hybridize to nascent RNA during the course of synthesis. We f...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gnh068

    authors: Marras SA,Gold B,Kramer FR,Smith I,Tyagi S

    更新日期:2004-05-20 00:00:00

  • DNA3'pp5'G de-capping activity of aprataxin: effect of cap nucleoside analogs and structural basis for guanosine recognition.

    abstract::DNA3'pp5'G caps synthesized by the 3'-PO4/5'-OH ligase RtcB have a strong impact on enzymatic reactions at DNA 3'-OH ends. Aprataxin, an enzyme that repairs A5'pp5'DNA ends formed during abortive ligation by classic 3'-OH/5'-PO4 ligases, is also a DNA 3' de-capping enzyme, converting DNAppG to DNA3'p and GMP. By takin...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv501

    authors: Chauleau M,Jacewicz A,Shuman S

    更新日期:2015-07-13 00:00:00

  • Mouse Genome Database (MGD)-2017: community knowledge resource for the laboratory mouse.

    abstract::The Mouse Genome Database (MGD: http://www.informatics.jax.org) is the primary community data resource for the laboratory mouse. It provides a highly integrated and highly curated system offering a comprehensive view of current knowledge about mouse genes, genetic markers and genomic features as well as the associatio...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw1040

    authors: Blake JA,Eppig JT,Kadin JA,Richardson JE,Smith CL,Bult CJ,the Mouse Genome Database Group.

    更新日期:2017-01-04 00:00:00

  • Competitive inactivation of a double-strand DNA break site involves parallel suppression of meiosis-induced changes in chromatin configuration.

    abstract::In Saccharomyces cerevisiae, DNA double-strand breaks (DSBs) initiate meiotic recombination at open sites in chromatin, which display a meiosis-specific increase in micrococcal nuclease (MNase) sensitivity. The arg4 promoter contains such a DSB site. When arg4 sequences are placed in a pBR322-derived insert at HIS4 (h...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/27.10.2175

    authors: Ohta K,Wu TC,Lichten M,Shibata T

    更新日期:1999-05-15 00:00:00

  • MyHits: improvements to an interactive resource for analyzing protein sequences.

    abstract::The MyHits web site (http://myhits.isb-sib.ch) is an integrated service dedicated to the analysis of protein sequences. Since its first description in 2004, both the user interface and the back end of the server were improved. A number of tools (e.g. MAFFT, Jacop, Dotlet, Jalview, ESTScan) were added or updated to imp...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm352

    authors: Pagni M,Ioannidis V,Cerutti L,Zahn-Zabal M,Jongeneel CV,Hau J,Martin O,Kuznetsov D,Falquet L

    更新日期:2007-07-01 00:00:00

  • Tissue-specific transcript annotation and expression profiling with complementary next-generation sequencing technologies.

    abstract::Next-generation sequencing is excellently suited to evaluate the abundance of mRNAs to study gene expression. Here we compare two alternative technologies, cap analysis of gene expression (CAGE) and serial analysis of gene expression (SAGE), for the same RNA samples. Along with quantifying gene expression levels, CAGE...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq602

    authors: Hestand MS,Klingenhoff A,Scherf M,Ariyurek Y,Ramos Y,van Workum W,Suzuki M,Werner T,van Ommen GJ,den Dunnen JT,Harbers M,'t Hoen PA

    更新日期:2010-09-01 00:00:00

  • The Universal Protein Resource (UniProt).

    abstract::The ability to store and interconnect all available information on proteins is crucial to modern biological research. Accordingly, the Universal Protein Resource (UniProt) plays an increasingly important role by providing a stable, comprehensive, freely accessible central resource on protein sequences and functional a...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl929

    authors: UniProt Consortium.

    更新日期:2007-01-01 00:00:00

  • PAH Mutation Analysis Consortium Database: 1997. Prototype for relational locus-specific mutation databases.

    abstract::PAHdb (http://www.mcgill.ca/pahdb ) is a curated relational database (Fig. 1) of nucleotide variation in the human PAH cDNA (GenBank U49897). Among 328 different mutations by state (Fig. 2) the majority are rare mutations causing hyperphenylalaninemia (HPA) (OMIM 261600), the remainder are polymorphic variants without...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/26.1.220

    authors: Nowacki PM,Byck S,Prevost L,Scriver CR

    更新日期:1998-01-01 00:00:00

  • DNA multiplex hybridization on microarrays and thermodynamic stability in solution: a direct comparison.

    abstract::Hybridization intensities of 30 distinct short duplex DNAs measured on spotted microarrays, were directly compared with thermodynamic stabilities measured in solution. DNA sequences were designed to promote formation of perfect match, or hybrid duplexes containing tandem mismatches. Thermodynamic parameters DeltaH deg...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm865

    authors: Fish DJ,Horne MT,Brewood GP,Goodarzi JP,Alemayehu S,Bhandiwad A,Searles RP,Benight AS

    更新日期:2007-01-01 00:00:00

  • Discovery and characterization of a thermostable bacteriophage RNA ligase homologous to T4 RNA ligase 1.

    abstract::Thermophilic viruses represent a novel source of genetic material and enzymes with great potential for use in biotechnology. We have isolated a number of thermophilic viruses from geothermal areas in Iceland, and by combining high throughput genome sequencing and state of the art bioinformatics we have identified a nu...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg914

    authors: Blondal T,Hjorleifsdottir SH,Fridjonsson OF,Aevarsson A,Skirnisdottir S,Hermannsdottir AG,Hreggvidsson GO,Smith AV,Kristjansson JK

    更新日期:2003-12-15 00:00:00

  • Assessment of clone identity and sequence fidelity for 1189 IMAGE cDNA clones.

    abstract::This report documents the error rate in a commercially distributed subset of the IMAGE Consortium mouse cDNA clone collection. After isolation of plasmid DNA from 1189 bacterial stock cultures, only 62. 2% were uncontaminated and contained cDNA inserts that had significant sequence identity to published data for the o...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.2.582

    authors: Halgren RG,Fielden MR,Fong CJ,Zacharewski TR

    更新日期:2001-01-15 00:00:00

  • Vitamin D receptor contains multiple dimerization interfaces that are functionally different.

    abstract::The vitamin D receptor mediates the signal of 1 alpha, 25-dihydroxyvitamin D3 by binding to vitamin D responsive elements in DNA as a homodimer or as a heterodimer composed of one vitamin D receptor subunit and one retinoid X receptor subunit. We have mapped the dimerization interfaces of the vitamin D receptor that i...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.4.606

    authors: Nishikawa J,Kitaura M,Imagawa M,Nishihara T

    更新日期:1995-02-25 00:00:00

  • U1-independent pre-mRNA splicing contributes to the regulation of alternative splicing.

    abstract::U1 snRNP plays a crucial role in the 5' splice site recognition during splicing. Here we report the first example of naturally occurring U1-independent U2-type splicing in humans. The U1 components were not included in the pre-spliceosomal E complex formed on the human F1gamma (hF1gamma) intron 9 in vitro. Moreover, h...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp050

    authors: Fukumura K,Taniguchi I,Sakamoto H,Ohno M,Inoue K

    更新日期:2009-04-01 00:00:00

  • An on-bead tailing/ligation approach for sequencing resin-bound RNA libraries.

    abstract::Nucleic acids possess the unique property of being enzymatically amplifiable, and have therefore been a popular choice for the combinatorial selection of functional sequences, such as aptamers or ribozymes. However, amplification typically requires known sequence segments that serve as primer binding sites, which can ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks004

    authors: Wiesmayr A,Fournier P,Jäschke A

    更新日期:2012-05-01 00:00:00

  • Early steps of active DNA demethylation initiated by ROS1 glycosylase require three putative helix-invading residues.

    abstract::Active DNA demethylation is crucial for epigenetic control, but the underlying enzymatic mechanisms are incompletely understood. REPRESSOR OF SILENCING 1 (ROS1) is a 5-methylcytosine (5-meC) DNA glycosylase/lyase that initiates DNA demethylation in plants through a base excision repair process. The enzyme binds DNA no...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkt625

    authors: Parrilla-Doblas JT,Ponferrada-Marín MI,Roldán-Arjona T,Ariza RR

    更新日期:2013-10-01 00:00:00

  • Rolling circle amplification shows a sinusoidal template length-dependent amplification bias.

    abstract::Biophysical properties of DNA such as its longitudinal and torsional persistence length govern many processes and phenomena in biology, DNA nanotechnology and biotechnology. It has, for example, long been known that the circularization efficiency of short DNA fragments shows a periodic pattern where fragments with int...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx1238

    authors: Joffroy B,Uca YO,Prešern D,Doye JPK,Schmidt TL

    更新日期:2018-01-25 00:00:00

  • Expanded molecular diversity generation during directed evolution by trinucleotide exchange (TriNEx).

    abstract::Trinucleotide exchange (TriNEx) is a method for generating novel molecular diversity during directed evolution by random substitution of one contiguous trinucleotide sequence for another. Single trinucleotide sequences were deleted at random positions in a target gene using the engineered transposon MuDel that were su...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkn358

    authors: Baldwin AJ,Busse K,Simm AM,Jones DD

    更新日期:2008-08-01 00:00:00

  • RNA polymerase III dependence of the human L1 promoter and possible participation of the RNA polymerase II factor YY1 in the RNA polymerase III transcription system.

    abstract::From the general views of the eukaryotic transcription systems, L1 (or L1-like) retrotransposons that encode some proteins are unusual. L1, unlike other protein-coding elements, is transcribed through an internal promoter. And the L1 internal promoter, unlike other internal promoters, is thought to be RNA polymerase I...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.18.3704

    authors: Kurose K,Hata K,Hattori M,Sakaki Y

    更新日期:1995-09-25 00:00:00

  • Synthesis, structure and thermodynamic properties of 8-methylguanine-containing oligonucleotides: Z-DNA under physiological salt conditions.

    abstract::Various oligonucleotides containing 8-methylguanine (m8G) have been synthesized and their structures and thermodynamic properties investigated. Introduction Of M8G into DNA sequences markedly stabilizes the Z conformation under low salt conditions. The hexamer d(CGC[M8G]CG)2 exhibits a CD spectrum characteristic of th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/24.7.1272

    authors: Sugiyama H,Kawai K,Matsunaga A,Fujimoto K,Saito I,Robinson H,Wang AH

    更新日期:1996-04-01 00:00:00

  • APC gene: database of germline and somatic mutations in human tumors and cell lines.

    abstract::A database (http://perso.curie.fr/tsoussi ) is described, in which over 1000 mutations in the human APC gene of tumors (colon cancer predominantly) are compiled from the literature. It includes both molecular information about the mutations and clinical data about the patients. Software has been designed to analyse al...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/26.1.269

    authors: Laurent-Puig P,Béroud C,Soussi T

    更新日期:1998-01-01 00:00:00

  • Crosstalk between transposase subunits during cleavage of the mariner transposon.

    abstract::Mariner transposition is a complex reaction that involves three recombination sites and six strand breaking and joining reactions. This requires precise spatial and temporal coordination between the different components to ensure a productive outcome and minimize genomic instability. We have investigated how the cleav...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku172

    authors: Claeys Bouuaert C,Walker N,Liu D,Chalmers R

    更新日期:2014-05-01 00:00:00

  • Covalent attachment of synthetic DNA to self-assembled monolayer films.

    abstract::The covalent attachment of thiol-modified DNA oligomers; to self-assembled monolayer silane films on fused silica and oxidized silicon substrates is described. A heterobifunctional crosslinking molecule bearing both thiol- and amino-reactive moieties was used to tether a DNA oligomer (modified at its terminus with a t...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/24.15.3031

    authors: Chrisey LA,Lee GU,O'Ferrall CE

    更新日期:1996-08-01 00:00:00

  • Experimental maps of DNA structure at nucleotide resolution distinguish intrinsic from protein-induced DNA deformations.

    abstract::Recognition of DNA by proteins depends on DNA sequence and structure. Often unanswered is whether the structure of naked DNA persists in a protein-DNA complex, or whether protein binding changes DNA shape. While X-ray structures of protein-DNA complexes are numerous, the structure of naked cognate DNA is seldom availa...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gky033

    authors: Azad RN,Zafiropoulos D,Ober D,Jiang Y,Chiu TP,Sagendorf JM,Rohs R,Tullius TD

    更新日期:2018-03-16 00:00:00

  • Evidence that RNA editing modulates splice site selection in the 5-HT2C receptor gene.

    abstract::Adenosine to inosine editing of mRNA from the human 5-HT2C receptor gene (HTR2C) occurs at five exonic positions (A-E) in a stable stem-loop that includes the normal 5' splice site of intron 5 and is flanked by two alternative splice sites. Using in vitro editing, we identified a novel editing site (F) located in the ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkh536

    authors: Flomen R,Knight J,Sham P,Kerwin R,Makoff A

    更新日期:2004-04-15 00:00:00