Abstract:
:Antisense oligodeoxynucleotides targeted to Ha-ras mRNA have been designed to discriminate between the codon 12-mutated oncogene and the normal proto-oncogene. An in vitro assay using two different sources of RNase H (rabbit reticulocyte lysates and nuclear extract from HeLa cells) was used to characterize oligonucleotide binding to normal and mutated Ha-ras mRNA. Short oligonucleotides (12- or 13mers) centered on the mutation had a very high discriminatory efficiency. Longer oligonucleotides (16mers) did not discriminate efficiently between the mutated and the normal mRNA. We have tested the efficacy of dodecanucleotides to induce RNase H cleavage of the full-length mRNA, moving the target sequence from the loop to the stem region which is formed in the vicinity of mutated codon 12. The most selective oligonucleotides were centered on the mutation which is located near the junction between the loop and stem regions even though they were less efficient at inducing RNase H cleavage than those targeted to the loop region. The 12mer antisense oligonucleotide with the highest discriminatory power was selected for cell culture studies. This oligonucleotide inhibited the proliferation of a human cell line which had been transformed with the mutated Ha-ras gene (HBL100ras1) but had no effect on the parental cell line which was transfected with the vector DNA (HBL 100neo) and expressed only the normal Ha-ras gene. Growth inhibition of HBL100ras1 cells was associated with specific ablation of targeted Ha-ras mRNA as shown by RT-PCR. These results show that 'in vitro' evaluation using an RNase H assay allowed us to select an antisense oligonucleotide which elicited a selectivity towards point-mutated Ha-ras mRNA when added at 10 microM concentration to the culture medium of cells expressing wild type and mutated Ha-ras mRNA.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Duroux I,Godard G,Boidot-Forget M,Schwab G,Hélène C,Saison-Behmoaras Tdoi
10.1093/nar/23.17.3411subject
Has Abstractpub_date
1995-09-11 00:00:00pages
3411-8issue
17eissn
0305-1048issn
1362-4962pii
5c0122journal_volume
23pub_type
杂志文章abstract::The E.coli ada gene protein coding region has been ligated into an extrachromosomally replicating yeast expression vector downstream of the yeast alcohol dehydrogenase gene promoter region to produce pADH06C. The yeast strains SX46A, 7799-4B and VV-6 are deficient in endogenous O6-alkylguanine-DNA-alkyltransferase and...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.2.331
更新日期:1990-01-25 00:00:00
abstract::A new integrated image analysis package with quantitative quality control schemes is described for cDNA microarray technology. The package employs an iterative algorithm that utilizes both intensity characteristics and spatial information of the spots on a microarray image for signal-background segmentation and define...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.15.e75
更新日期:2001-08-01 00:00:00
abstract::DNA polymerase delta (Pol δ) is responsible for the elongation and maturation of Okazaki fragments in eukaryotic cells. Proliferating cell nuclear antigen (PCNA) recruits Pol δ to the DNA and serves as a processivity factor. Here, we show that PCNA also stimulates the catalytic rate of Saccharomyces cerevisiae Pol δ b...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky1321
更新日期:2019-02-28 00:00:00
abstract::Alternative splicing (AS) and alternative polyadenylation (APA) generate diverse transcripts in mammalian genomes during development and differentiation. Epigenetic marks such as trimethylation of histone H3 lysine 36 (H3K36me3) and DNA methylation play a role in generating transcriptome diversity. Intragenic CpG isla...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa556
更新日期:2020-09-04 00:00:00
abstract::The major mRNA degradation pathway involves deadenylation of the target molecule followed by decapping and, finally, 5'-->3' exonuclease digestion of the mRNA body. While yeast factors involved in the decapping and exonuclease degradation steps have been identified, the nature of the factor(s) involved in the deadenyl...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.12.2448
更新日期:2001-06-15 00:00:00
abstract::Members of several classes of proto-oncogenes replicate during the first third of S-phase in two human (K562 erythroleukemia and HeLa), one Chinese hamster (CHO) and eight mouse cell lines. These cell lines exhibit a variety of specialized functions characteristic of pre-B and B cells, T cells and erythroid cells. The...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.1.87
更新日期:1987-01-12 00:00:00
abstract::Diazo-based precursors of photolabile groups have been used extensively for modifying nucleic acids, with the intention of toggling biological processes with light. These processes include transcription, translation and RNA interference. In these cases, the photolabile groups have been typically depicted as modifying ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp415
更新日期:2009-07-01 00:00:00
abstract::We have prepared several novel phosphoramidites and have synthesised oligonucleotides incorporating them internally. The presence of these residues in an oligonucleotide template presents an impossible barrier to primed synthesis by Taq DNA polymerase. When extended as polymerase chain reaction products, these oligonu...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.5.1155
更新日期:1993-03-11 00:00:00
abstract::The Ensembl project has been aggregating, processing, integrating and redistributing genomic datasets since the initial releases of the draft human genome, with the aim of accelerating genomics research through rapid open distribution of public data. Large amounts of raw data are thus transformed into knowledge, which...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx1098
更新日期:2018-01-04 00:00:00
abstract::Regulation of gene expression influences almost all biological processes in an organism; sequence-specific DNA-binding transcription factors are critical to this control. For most genomes, the repertoire of transcription factors is only partially known. Hitherto transcription factor identification has been largely bas...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkj131
更新日期:2006-01-01 00:00:00
abstract::Topoisomerases (topos) maintain DNA topology and influence DNA transaction processes by catalysing relaxation, supercoiling and decatenation reactions. In the cellular milieu, division of labour between different topos ensures topological homeostasis and control of central processes. In Escherichia coli, DNA gyrase is...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks843
更新日期:2012-11-01 00:00:00
abstract::Analysis of large-scale gene expression studies usually begins with gene clustering. A ubiquitous problem is that different algorithms applied to the same data inevitably give different results, and the differences are often substantial, involving a quarter or more of the genes analyzed. This raises a series of import...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki536
更新日期:2005-05-10 00:00:00
abstract::The Xist lncRNA requires Repeat A, a conserved RNA element located in its 5' end, to induce gene silencing during X-chromosome inactivation. Intriguingly, Repeat A is also required for production of Xist. While silencing by Repeat A requires the protein SPEN, how Repeat A promotes Xist production remains unclear. We r...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa789
更新日期:2020-10-09 00:00:00
abstract::Human Ape2 protein has 3' phosphodiesterase activity for processing 3'-damaged DNA termini, 3'-5' exonuclease activity that supports removal of mismatched nucleotides from the 3'-end of DNA, and a somewhat weak AP-endonuclease activity. However, very little is known about the role of Ape2 in DNA repair processes. Here...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkp357
更新日期:2009-07-01 00:00:00
abstract::With the high number of sequences and structures streaming in from genomic projects, there is a need for more powerful and sophisticated annotation tools. Most problematic of the annotation efforts is predicting gene and protein function. Over the past few years there has been considerable progress in automated protei...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl045
更新日期:2006-07-01 00:00:00
abstract::Nuclear organization of chromatin is an important level of genome regulation with positional changes of genes occurring during reprogramming. Inherent variability of biological specimens, wide variety of sample preparation and imaging conditions, though pose significant challenges to data analysis and comparison. Here...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr486
更新日期:2011-09-01 00:00:00
abstract::RNase P is primarily responsible for the 5΄ maturation of transfer RNAs (tRNAs) in all domains of life. Archaeal RNase P is a ribonucleoprotein made up of one catalytic RNA and five protein cofactors including L7Ae, which is known to bind the kink-turn (K-turn), an RNA structural element that causes axial bending. How...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx388
更新日期:2017-07-07 00:00:00
abstract::A fundamental problem with applying Consensus, Weight-Matrix or hidden Markov models as search tools for biosequences is that there is no way to know, from the model, if the modeled sequences display any dependencies between positional identities. In some instances, these dependencies are crucial in correctly acceptin...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gki374
更新日期:2005-07-01 00:00:00
abstract::Arginyl-tRNA synthetase (ArgRS) catalyses AMP- and PPi-independent deacylation of Arg-tRNAArg in the presence of cysteine. A dipeptide, Arg-Cys, is a product of this deacylation reaction. Similar reaction with homocysteine yields Arg-Hcy. Arginine is a noncompetitive inhibitor of the cysteine-dependent deacylation whi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/23.22.4608
更新日期:1995-11-25 00:00:00
abstract::Scanning tunneling microscopy (STM) reveals nanometer scale details of hydrated DNA but the interpretation of the images is controversial because of substrate artifacts and the lack of a theory for image contrast. We demonstrate that we have overcome these problems by identifying five DNA samples by their STM images a...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.25.5896
更新日期:1993-12-25 00:00:00
abstract::A general procedure, based on a new activated alkyne linker, for the preparation of peptide-oligonucleotide conjugates (POCs) on solid support has been developed. With this linker, conjugation is effective at room temperature (RT) in millimolar concentration and submicromolar amounts. This is made possible since the u...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr603
更新日期:2011-11-01 00:00:00
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
更新日期:2012-05-01 00:00:00
abstract::The MF alpha 1 gene of Saccharomyces cerevisiae is an alpha specific gene whose expression is regulated by two upstream activation sites (UASMF alpha 1S), which are the binding sites for activator proteins, MAT alpha 1 and MCM1. UASMF alpha 1 elements act in a synergistic fashion; lack of either element causes reduced...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/19.11.3099
更新日期:1991-06-11 00:00:00
abstract::The accumulation of somatic mitochondrial DNA (mtDNA) mutations contributes to the pathogenesis of human disease. Currently, mitochondrial mutations are largely considered results of inaccurate processing of its heavily damaged genome. However, mainly from a lack of methods to monitor mtDNA mutations with sufficient s...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw716
更新日期:2016-10-14 00:00:00
abstract::The G- and U-specific reagent, carbodiimide was used to probe the solution structure of aminoacylated yeast phenylalanine tRNA. Both quantitative and qualitative changes in modification were observed when the modification patterns of tRNA-CCA(3'OH), tRNA-CCA(3'NH2) and phe-tRNA-CCA(3'NH2) were compared. Five nucleotid...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.7.2419
更新日期:1982-04-10 00:00:00
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::A RNA interference (RNAi) like mechanism is involved in elimination of thousands of DNA segments from the developing somatic macronucleus of Tetrahymena, yet how specific internal eliminated sequences (IESs) are recognized remains to be fully elucidated. To define requirements for DNA rearrangement, we performed mutag...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl699
更新日期:2006-01-01 00:00:00
abstract::An alpha-neo-endorphin (alpha NE) gene, which we previously synthesized chemically and inserted into E. coli beta-galactosidase gene of pK013 plasmid, has been excised and fused to E. coli alkaline phosphatase (APase) gene. One of the transformants was named E15/pA alpha NE1. Under the APase gene regulation, APase-alp...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/11.5.1283
更新日期:1983-03-11 00:00:00
abstract::We have isolated a unique fragment of the HLA-DR alpha gene and probed human genomic DNA at low stringency to search for homologous sequences. A minimum of six non-polymorphic cross-hybridizing high molecular weight fragments were found in all DNAs examined. In order to obtain molecular clones of these cross-hybridizi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/13.20.7515
更新日期:1985-10-25 00:00:00
abstract::We characterize a component of the E. coli bacterial nucleoid H1a, which accumulates in stationary phase. This protein, identical with the major component of a plasmid-protein complex previously isolated in our laboratory, has a pI close to 7.5. Acrylamide gel electrophoresis and sedimentation in sucrose gradient have...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.13.5321
更新日期:1984-07-11 00:00:00