Abstract:
:Several mono-, di-, tetra-, penta- and nonaribozymes were developed. These multitarget-ribozymes were targeted to cleave HIV-1 env RNA at up to nine different conserved sites. Each multitarget-ribozyme consisted of a chain of up to nine hammerhead motifs, each flanked by a different targeting sequence. The multitarget-ribozymes were functional in vitro and gave rise to multiple, specific partial and/or complete RNA digestion products. Per RNA copy, multitarget-ribozymes were more efficient than monoribozymes or ribozymes targeting a subset of the same sites. In contrast to monoribozymes, a 400nt nonaribozyme, targeted to cleave at nine different sites within a 1.3kb HIV-1 env RNA substrate, was active and showed the same specificity of cleavage when it was part of a large 3.3kb transcript. We conclude that multitarget-ribozymes retain the specificity of monoribozymes, but they are more efficient per ribozyme RNA copy and they remain active when they are part of a large transcript. A tetra-, penta- or nonaribozyme under control of the SV40 late promoter, the beta-actin gene promoter or the HIV-1 LTR, respectively, were cotransfected with the infectious HIV-1 DNA clone pNL4-3 into permissive HeLa T4 cells. Each cotransfection resulted in a specific inhibition of HIV-1 replication as determined by syncytia formation and p24 antigen release. In addition, coexpression of the nonaribozyme with an HIV-1 env RNA transcript resulted in the specific dramatic reduction of the env transcript. We conclude that the multitarget-ribozymes are also functional intracellularly. A nucleotide sequence comparison of the target sites indicates that the multitarget-ribozymes could potentially be effective against all thirty HIV-1 isolates presently sequenced. Their use may help to slow the selection of viral escape mutants and thereby prolong their effectiveness. We anticipate that multitarget-ribozymes will also be more effective in the successful targeting of less variable cellular RNAs.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Chen CJ,Banerjea AC,Harmison GG,Haglund K,Schubert Mdoi
10.1093/nar/20.17.4581keywords:
subject
Has Abstractpub_date
1992-09-11 00:00:00pages
4581-9issue
17eissn
0305-1048issn
1362-4962journal_volume
20pub_type
杂志文章abstract::mRNA expression involves transcription initiation, elongation and degradation. In cells, these dynamic processes are highly regulated. However, experimental characterization of the dynamic processes in vivo is difficult due to the paucity of methods capable of direct measurements. We present a highly sensitive and ver...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw596
更新日期:2016-10-14 00:00:00
abstract::The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-2H]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60 degrees C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.19.5131
更新日期:1992-10-11 00:00:00
abstract::HAP1, also known as APE/Ref-1, is the major apurinic/apyrimidinic (AP) endonuclease in human cells. Previous structural studies have suggested a possible role for the Asp-210 residue of HAP1 in the enzymatic function of this enzyme. Here, we demonstrate that substitution of Asp-210 by Asn or Ala eliminates the AP endo...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.11.2207
更新日期:2000-06-01 00:00:00
abstract::A study of the synthesis of oligothymidylates via phosphotriester intermediates on a polystyrene support is described. The sequence involves condensation of a phenyl nucleoside-3' -phosphorochloridate with the 5'-hydroxyl group of the carrier bound oligonucleotide derivative in the presence of 1-methylimidazole. Condi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.6.773
更新日期:1975-06-01 00:00:00
abstract::In this study, we demonstrate that the lack of retinoic acid-related orphan receptor (ROR) γ or α expression in mice significantly reduced the peak expression level of Cry1, Bmal1, E4bp4, Rev-Erbα and Per2 in an ROR isotype- and tissue-selective manner without affecting the phase of their rhythmic expression. Analysis...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks630
更新日期:2012-09-01 00:00:00
abstract::Chlorella virus DNA ligase is the smallest eukaryotic ATP-dependent DNA ligase known; it suffices for yeast cell growth in lieu of the essential yeast DNA ligase Cdc9. The Chlorella virus ligase-adenylate intermediate has an intrinsic nick sensing function and its DNA footprint extends 8-9 nt on the 3'-hydroxyl (3'-OH...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg665
更新日期:2003-09-01 00:00:00
abstract::A cDNA (2855 nt) encoding a putative cytotoxic ribonuclease (rapLR1) related to the antitumor protein onconase was cloned from a library derived from the liver of gravid female amphibian Rana pipiens. The cDNA was mainly comprised (83%) of 3' untranslated region (UTR). Secondary structure analysis predicted two unusua...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.12.2375
更新日期:2000-06-15 00:00:00
abstract::As the formation of ribonucleoprotein complexes is a major mechanism of angiotensin II type 1 receptor (AT1R) regulation, we sought to identify novel AT1R mRNA binding proteins. By affinity purification and mass spectroscopy, we identified TIA-1. This interaction was confirmed by colocalization of AT1R mRNA and TIA-1 ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv1368
更新日期:2016-04-20 00:00:00
abstract::The widespread applications of various 'omics' technologies in biomedical research together with the emergence of public data repositories have resulted in a plethora of data sets for almost any given physiological state or disease condition. Properly combining or integrating these data sets with similar basic hypothe...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt338
更新日期:2013-07-01 00:00:00
abstract::In this article, we characterize histone demethylase activity of the entire family of JmjC+N proteins of Drosophila melanogaster. Our results show that Lid (little imaginal discs), which is structurally homologous to JARID1, demethylates H3K4me3. However, contrary to what would be inferred from its demethylase activit...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkn098
更新日期:2008-05-01 00:00:00
abstract::Tomato golden mosaic geminivirus has a genome of two single-stranded (ss) DNA components, A and B. An almost identical 'common' region in DNA A and DNA B is thought to contain sequence elements controlling replication and transcription. Hence investigation of sequences important for DNA replication by in vitro mutagen...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/17.24.10213
更新日期:1989-12-25 00:00:00
abstract::Guanine-quadruplexes (G4) consist of non-canonical four-stranded helical arrangements of guanine-rich nucleic acid sequences. The bulky and thermodynamically stable features of G4 structures have been shown in many respects to affect normal nucleic acid metabolism. In vivo conversion of G4 structures to single-strande...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr630
更新日期:2011-11-01 00:00:00
abstract::The development of genetic switches and their integrated forms (genetic circuits) with desired specifications/functions is key for success in synthetic biology. Due to the difficulty in rational design, genetic switches and circuits with desirable specifications are mostly obtained by directed evolution. Based on a vi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq1070
更新日期:2011-02-01 00:00:00
abstract::Point mutations in mitochondrial tRNA genes are responsible for individual subgroups of mitochondrial encephalomyopathies. We have recently reported that point mutations in the tRNA(Leu)(UUR) and tRNA(Lys) genes cause a defect in the normal modification at the first nucleotide of the anticodon. As part of a systematic...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.19.3779
更新日期:2000-10-01 00:00:00
abstract::Co-option of cis-regulatory modules has been suggested as a mechanism for the evolution of expression sites during development. However, the extent and mechanisms involved in mobilization of cis-regulatory modules remains elusive. To trace the history of non-coding elements, which may represent candidate ancestral cis...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt030
更新日期:2013-04-01 00:00:00
abstract::We have adapted a method for making libraries of mutations in any specific gene for use in the fission yeast Schizosaccharomyces pombe . This elegant and simple method consists of PCR amplification of the gene of interest, followed by co-transformation of fission yeast with the PCR fragment and a linearized plasmid ve...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.20.4783
更新日期:1998-10-15 00:00:00
abstract::enod40 is a plant gene that participates in the regulation of symbiotic interaction between leguminous plants and bacteria or fungi. Furthermore, it has been suggested to play a general role in non-symbiotic plant development. Although enod40 seems to have multiple functions, being present in many land plants, the mol...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm173
更新日期:2007-01-01 00:00:00
abstract::The centromere is a highly specialized chromosomal element that is essential for chromosome segregation during mitosis. Centromere integrity must therefore be properly preserved and is strictly dependent upon the establishment and maintenance of surrounding chromatin structure. Here we identify WDHD1, a WD40-domain an...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq1338
更新日期:2011-05-01 00:00:00
abstract::Plasmid encoded replication initiation (Rep) proteins recruit host helicases to plasmid replication origins. Previously, we showed that RepD recruits directionally the PcrA helicase to the pC221 oriD, remains associated with it, and increases its processivity during plasmid unwinding. Here we show that RepD forms a co...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp1153
更新日期:2010-04-01 00:00:00
abstract::The Ribosomal Database Project (RDP-II) provides the research community with aligned and annotated rRNA gene sequences, along with analysis services and a phylogenetically consistent taxonomic framework for these data. Updated monthly, these services are made available through the RDP-II website (http://rdp.cme.msu.ed...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki038
更新日期:2005-01-01 00:00:00
abstract::Genomic DNA of African trypanosomes contains a hypermodified thymidine residue termed base J (beta-d-glucosyl-HOMedU). This modified base is localized primarily to repetitive DNA, namely the telomeres, and is implicated in the regulation of antigenic variation. The base is synthesized in a two-step pathway. Initially,...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkn1067
更新日期:2009-04-01 00:00:00
abstract::Initiation of bacterial DNA replication at oriC is mediated by primosomal proteins that act cooperatively to melt an AT-rich region where the replicative helicase is loaded prior to the assembly of the replication fork. In Bacillus subtilis, the dnaD, dnaB and dnaI genes are essential for initiation of DNA replication...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp1236
更新日期:2010-05-01 00:00:00
abstract::In non-malignant RWPE-1 prostate epithelial cells signaling by the nuclear receptor Vitamin D Receptor (VDR, NR1I1) induces cell cycle arrest through targets including CDKN1A (encodes p21((waf1/cip1))). VDR dynamically induced individual histone modification patterns at three VDR binding sites (R1, 2, 3) on the CDKN1A...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq875
更新日期:2011-03-01 00:00:00
abstract::The Network of Cancer Genes (NCG) collects and integrates data on 736 human genes that are mutated in various types of cancer. For each gene, NCG provides information on duplicability, orthology, evolutionary appearance and topological properties of the encoded protein in a comprehensive version of the human protein-p...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp957
更新日期:2010-01-01 00:00:00
abstract::Chromatin assembly factor 1 (CAF-1) is a histone H3-H4 chaperone that deposits newly synthesized histone (H3-H4)2 tetramers during replication-coupled nucleosome assembly. However, how CAF-1 functions in this process is not yet well understood. Here, we report the crystal structure of C terminus of Cac1 (Cac1C), a sub...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw106
更新日期:2016-06-20 00:00:00
abstract::Oligo(dG).oligo(dC)- or short poly(dG).poly(dC)-containing fragments were enriched and cloned by means of Mg2+-dependent triplex affinity capture and subsequent cloning procedures. A library constructed after three cycles of enrichment showed that approximately 80% of the clones in the supercoiled form formed a comple...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/25.9.1701
更新日期:1997-05-01 00:00:00
abstract::Denaturing gradient gel electrophoresis (DGGE) is based upon the different melting behaviors of DNA molecules in a chemical denaturant gradient according to their sequences. This technique has recently become a widespread tool to detect mutations. The introduction of a GC-clamp enables the detection of most single bas...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.12.3331
更新日期:1991-06-25 00:00:00
abstract::Biological networks are a topic of great current interest, particularly with the publication of a number of large genome-wide interaction datasets. They are globally characterized by a variety of graph-theoretic statistics, such as the degree distribution, clustering coefficient, characteristic path length and diamete...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh164
更新日期:2004-01-14 00:00:00
abstract::IntAct is an open-source, open data molecular interaction database populated by data either curated from the literature or from direct data depositions. Two levels of curation are now available within the database, with both IMEx-level annotation and less detailed MIMIx-compatible entries currently supported. As from ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr1088
更新日期:2012-01-01 00:00:00
abstract::The archaeal and eukaryotic evolutionary domains diverged from each other approximately 2 billion years ago, but many of the core components of their transcriptional and translational machineries still display a readily recognizable degree of similarity in their primary structures. The F and P subunits present in arch...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.21.4299
更新日期:2000-11-01 00:00:00