Efficient production of superior dumbbell-shaped DNA minimal vectors for small hairpin RNA expression.

Abstract:

:Genetic therapy holds great promise for the treatment of inherited or acquired genetic diseases; however, its breakthrough is hampered by the lack of suitable gene delivery systems. Dumbbell-shaped DNA minimal vectors represent an attractive, safe alternative to the commonly used viral vectors which are fraught with risk, but dumbbell generation appears to be costly and time-consuming. We developed a new PCR-based method for dumbbell production which comprises only two steps. First, PCR amplification of the therapeutic expression cassette using chemically modified primers to form a ready-to-ligate DNA structure; and second, a highly efficient intramolecular ligation reaction. Compared with conventional strategies, the new method produces dumbbell vectors more rapidly, with higher yields and purity, and at lower costs. In addition, such produced small hairpin RNA expressing dumbbells triggered superior target gene knockdown compared with conventionally produced dumbbells or plasmids. Our novel method is suitable for large-scale dumbbell production and can facilitate clinical applications of this vector system.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Yu H,Jiang X,Tan KT,Hang L,Patzel V

doi

10.1093/nar/gkv583

subject

Has Abstract

pub_date

2015-10-15 00:00:00

pages

e120

issue

18

eissn

0305-1048

issn

1362-4962

pii

gkv583

journal_volume

43

pub_type

杂志文章
  • A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting.

    abstract::Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the resp...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/24.18.3653

    authors: Kobayashi K,Ohye T,Pastan I,Nagatsu T

    更新日期:1996-09-15 00:00:00

  • Structural dynamics of propeller loop: towards folding of RNA G-quadruplex.

    abstract::We have carried out an extended set of standard and enhanced-sampling MD simulations (for a cumulative simulation time of 620 μs) with the aim to study folding landscapes of the rGGGUUAGGG and rGGGAGGG parallel G-hairpins (PH) with propeller loop. We identify folding and unfolding pathways of the PH, which is bridged ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gky712

    authors: Havrila M,Stadlbauer P,Kührová P,Banáš P,Mergny JL,Otyepka M,Šponer J

    更新日期:2018-09-28 00:00:00

  • Genome-wide evidence for local DNA methylation spreading from small RNA-targeted sequences in Arabidopsis.

    abstract::Transposable elements (TEs) and their relics play major roles in genome evolution. However, mobilization of TEs is usually deleterious and strongly repressed. In plants and mammals, this repression is typically associated with DNA methylation, but the relationship between this epigenetic mark and TE sequences has not ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr324

    authors: Ahmed I,Sarazin A,Bowler C,Colot V,Quesneville H

    更新日期:2011-09-01 00:00:00

  • Transcription through the yeast origin of replication ARS1 ends at the ABFI binding site and affects extrachromosomal maintenance of minichromosomes.

    abstract::When the function of origins of replication in yeast was compromised by placing ARS sequences downstream of strong promoters, ARS activity might have been affected either by transcription or by an altered chromatin configuration induced by the construct. To distinguish between these possibilities, derivatives of the y...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.19.3904

    authors: Tanaka S,Halter D,Livingstone-Zatchej M,Reszel B,Thoma F

    更新日期:1994-09-25 00:00:00

  • Database for mobile group II introns.

    abstract::Group II introns are self-splicing RNAs and retroelements found in bacteria and lower eukaryotic organelles. During the past several years, they have been uncovered in surprising numbers in bacteria due to the genome sequencing projects; however, most of the newly sequenced introns are not correctly identified. We hav...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg049

    authors: Dai L,Toor N,Olson R,Keeping A,Zimmerly S

    更新日期:2003-01-01 00:00:00

  • Bhageerath: an energy based web enabled computer software suite for limiting the search space of tertiary structures of small globular proteins.

    abstract::We describe here an energy based computer software suite for narrowing down the search space of tertiary structures of small globular proteins. The protocol comprises eight different computational modules that form an automated pipeline. It combines physics based potentials with biophysical filters to arrive at 10 pla...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl789

    authors: Jayaram B,Bhushan K,Shenoy SR,Narang P,Bose S,Agrawal P,Sahu D,Pandey V

    更新日期:2006-01-01 00:00:00

  • Rapid shotgun cloning utilizing the two base recognition endonuclease CviJI.

    abstract::A new approach has been developed for the rapid fragmentation and fractionation of DNA into a size suitable for shotgun cloning and sequencing. The restriction endonuclease CviJI normally cleaves the recognition sequence PuGCPy between the G and C to leave blunt ends. Atypical reaction conditions which alter the speci...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/20.14.3753

    authors: Fitzgerald MC,Skowron P,Van Etten JL,Smith LM,Mead DA

    更新日期:1992-07-25 00:00:00

  • Predicting internal exons by oligonucleotide composition and discriminant analysis of spliceable open reading frames.

    abstract::A new method which predicts internal exon sequences in human DNA has been developed. The method is based on a splice site prediction algorithm that uses the linear discriminant function to combine information about significant triplet frequencies of various functional parts of splice site regions and preferences of ol...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.24.5156

    authors: Solovyev VV,Salamov AA,Lawrence CB

    更新日期:1994-12-11 00:00:00

  • Cell type-specific genomics of Drosophila neurons.

    abstract::Many tools are available to analyse genomes but are often challenging to use in a cell type-specific context. We have developed a method similar to the isolation of nuclei tagged in a specific cell type (INTACT) technique [Deal,R.B. and Henikoff,S. (2010) A simple method for gene expression and chromatin profiling of ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks671

    authors: Henry GL,Davis FP,Picard S,Eddy SR

    更新日期:2012-10-01 00:00:00

  • Bacteriophage lambda and plasmid vectors, allowing fusion of cloned genes in each of the three translational phases.

    abstract::We have constructed vectors from bacteriophage lambda and from plasmid pBR322 having a single EcoRI restriction site which is immediately downstream from the lac UV5 promotor. Each vector allows the fusion of a cloned gene to the lac Z gene in a different phase relative to the translation initiation codon of the lac Z...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/5.12.4479

    authors: Charnay P,Perricaudet M,Galibert F,Tiollais P

    更新日期:1978-12-01 00:00:00

  • Base specificity in the interaction of ethidium with synthetic polyribonucleotides.

    abstract::Base specificity in the interaction of ethidium with double stranded synthetic RNA homopolymers has been studied by means of spectroscopic (UV-visible absorption and fluorescence), microcalorimetric and dilatometric techniques. The results show a strong base specificity in this interaction, the association constant wi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/15.14.5803

    authors: Babayan Y,Manzini G,Xodo LE,Quadrifoglio F

    更新日期:1987-07-24 00:00:00

  • Position-dependent effects of regioisomeric methylated adenine and guanine ribonucleosides on translation.

    abstract::Reversible methylation of the N6 or N1 position of adenine in RNA has recently been shown to play significant roles in regulating the functions of RNA. RNA can also be alkylated upon exposure to endogenous and exogenous alkylating agents. Here we examined how regio-specific methylation at the hydrogen bonding edge of ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx515

    authors: You C,Dai X,Wang Y

    更新日期:2017-09-06 00:00:00

  • PCR performance of the B-type DNA polymerase from the thermophilic euryarchaeon Thermococcus aggregans improved by mutations in the Y-GG/A motif.

    abstract::The effect of mutations in the highly conserved Y-GG/A motif of B-type DNA polymerases was studied in the DNA polymerase from the hyperthermophilic euryarchaeon Thermococcus aggregans. This motif plays a critical role in the balance between the synthesis and degradation of the DNA chain. Five different mutations of th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/28.20.3910

    authors: Böhlke K,Pisani FM,Vorgias CE,Frey B,Sobek H,Rossi M,Antranikian G

    更新日期:2000-10-15 00:00:00

  • Atomistic insight into the kinetic pathways for Watson-Crick to Hoogsteen transitions in DNA.

    abstract::DNA predominantly contains Watson-Crick (WC) base pairs, but a non-negligible fraction of base pairs are in the Hoogsteen (HG) hydrogen bonding motif at any time. In HG, the purine is rotated ∼180° relative to the WC motif. The transitions between WC and HG may play a role in recognition and replication, but are diffi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz837

    authors: Vreede J,Pérez de Alba Ortíz A,Bolhuis PG,Swenson DWH

    更新日期:2019-12-02 00:00:00

  • DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.

    abstract::Type II CRISPR-Cas9 RNA-guided nucleases are widely used for genome engineering. Type II-A SpCas9 protein from Streptococcus pyogenes is the most investigated and highly used enzyme of its class. Nevertheless, it has some drawbacks, including a relatively big size, imperfect specificity and restriction to DNA targets ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz1225

    authors: Fedorova I,Arseniev A,Selkova P,Pobegalov G,Goryanin I,Vasileva A,Musharova O,Abramova M,Kazalov M,Zyubko T,Artamonova T,Artamonova D,Shmakov S,Khodorkovskii M,Severinov K

    更新日期:2020-02-28 00:00:00

  • Proteotoxic stress promotes entrapment of ribosomes and misfolded proteins in a shared cytosolic compartment.

    abstract::Cells continuously monitor protein synthesis to prevent accumulation of aberrant polypeptides. Insufficient capacity of cellular degradative systems, chaperone shortage or high levels of mistranslation by ribosomes can result in proteotoxic stress and endanger proteostasis. One of the least explored reasons for mistra...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa068

    authors: Ghosh A,Williams LD,Pestov DG,Shcherbik N

    更新日期:2020-04-17 00:00:00

  • ChimeRScope: a novel alignment-free algorithm for fusion transcript prediction using paired-end RNA-Seq data.

    abstract::The RNA-Seq technology has revolutionized transcriptome characterization not only by accurately quantifying gene expression, but also by the identification of novel transcripts like chimeric fusion transcripts. The 'fusion' or 'chimeric' transcripts have improved the diagnosis and prognosis of several tumors, and have...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx315

    authors: Li Y,Heavican TB,Vellichirammal NN,Iqbal J,Guda C

    更新日期:2017-07-27 00:00:00

  • Hierarchy of nonhomologous end-joining, single-strand annealing and gene conversion at site-directed DNA double-strand breaks.

    abstract::In mammalian cells, DNA double-strand breaks (DSBs) are repaired by three pathways, nonhomologous end-joining (NHEJ), gene conversion (GC) and single-strand annealing (SSA). These pathways are distinct with regard to repair efficiency and mutagenic potential and must be tightly controlled to preserve viability and gen...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkn347

    authors: Mansour WY,Schumacher S,Rosskopf R,Rhein T,Schmidt-Petersen F,Gatzemeier F,Haag F,Borgmann K,Willers H,Dahm-Daphi J

    更新日期:2008-07-01 00:00:00

  • scIGANs: single-cell RNA-seq imputation using generative adversarial networks.

    abstract::Single-cell RNA-sequencing (scRNA-seq) enables the characterization of transcriptomic profiles at the single-cell resolution with increasingly high throughput. However, it suffers from many sources of technical noises, including insufficient mRNA molecules that lead to excess false zero values, termed dropouts. Comput...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa506

    authors: Xu Y,Zhang Z,You L,Liu J,Fan Z,Zhou X

    更新日期:2020-09-04 00:00:00

  • CTC1-mediated C-strand fill-in is an essential step in telomere length maintenance.

    abstract::To prevent progressive telomere shortening as a result of conventional DNA replication, new telomeric DNA must be added onto the chromosome end. The de novo DNA synthesis involves elongation of the G-rich strand of the telomere by telomerase. In human cells, the CST complex (CTC1-STN1-TEN1) also functions in telomere ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx125

    authors: Feng X,Hsu SJ,Kasbek C,Chaiken M,Price CM

    更新日期:2017-05-05 00:00:00

  • Human N-ras: cDNA cloning and gene structure.

    abstract::The structure and organisation of the human N-ras gene has been determined by analysing cDNA clones derived from the two main mRNA transcripts. One clone in particular is 4.1 Kb long and originates from the larger (4.3 Kb) message. Sequence analysis of this clone has revealed that the N-ras gene consists of seven exon...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/13.14.5255

    authors: Hall A,Brown R

    更新日期:1985-07-25 00:00:00

  • Combining structure probing data on RNA mutants with evolutionary information reveals RNA-binding interfaces.

    abstract::Systematic structure probing experiments (e.g. SHAPE) of RNA mutants such as the mutate-and-map (MaM) protocol give us a direct access into the genetic robustness of ncRNA structures. Comparative studies of homologous sequences provide a distinct, yet complementary, approach to analyze structural and functional proper...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw217

    authors: Reinharz V,Ponty Y,Waldispühl J

    更新日期:2016-06-20 00:00:00

  • Search for differences in post-transcriptional modification patterns of mitochondrial DNA-encoded wild-type and mutant human tRNALys and tRNALeu(UUR).

    abstract::Post-transcriptional modifications are characteristic features of tRNAs and have been shown in a number of cases to influence both their structural and functional properties, including structure stabilization, amino-acylation and codon recognition. We have developed an approach which allows the investigation of the po...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/27.3.756

    authors: Helm M,Florentz C,Chomyn A,Attardi G

    更新日期:1999-02-01 00:00:00

  • 'Genome design' model and multicellular complexity: golden middle.

    abstract::Human tissue-specific genes were reported to be longer than housekeeping genes (both in coding and intronic parts). The competing neutralist and adaptationist models were proposed to explain this observation. Here I show that in human genome the longest are genes with the intermediate expression pattern. From the stan...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl773

    authors: Vinogradov AE

    更新日期:2006-01-01 00:00:00

  • Fast optimal alignment.

    abstract::We show how to speed up sequence alignment algorithms of the type introduced by Needleman and Wunsch (and generalized by Sellers and others). Faster alignment algorithms have been introduced, but always at the cost of possibly getting sub-optimal alignments. Our modification results in the optimal alignment still bein...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.1part1.175

    authors: Fickett JW

    更新日期:1984-01-11 00:00:00

  • Structural rearrangements in mRNA upon its binding to human 80S ribosomes revealed by EPR spectroscopy.

    abstract::The model mRNA (MR), 11-mer RNA containing two nitroxide spin labels at the 5'- and 3'-terminal nucleotides and prone to form a stable homodimer (MR)2, was used for Electron Paramagnetic Resonance study of structural rearrangements in mRNA occurring upon its binding to human 80S ribosomes. The formation of two differe...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx1136

    authors: Malygin AA,Graifer DM,Meschaninova MI,Venyaminova AG,Timofeev IO,Kuzhelev AA,Krumkacheva OA,Fedin MV,Karpova GG,Bagryanskaya EG

    更新日期:2018-01-25 00:00:00

  • BCNTB bioinformatics: the next evolutionary step in the bioinformatics of breast cancer tissue banking.

    abstract::Here, we present an update of Breast Cancer Now Tissue Bank bioinformatics, a rich platform for the sharing, mining, integration and analysis of breast cancer data. Its modalities provide researchers with access to a centralised information gateway from which they can access a network of bioinformatic resources to que...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx913

    authors: Gadaleta E,Pirrò S,Dayem Ullah AZ,Marzec J,Chelala C

    更新日期:2018-01-04 00:00:00

  • Evolutionary Conserved Motif Finder (ECMFinder) for genome-wide identification of clustered YY1- and CTCF-binding sites.

    abstract::We have developed a new bioinformatics approach called ECMFinder (Evolutionary Conserved Motif Finder). This program searches for a given DNA motif within the entire genome of one species and uses the gene association information of a potential transcription factor-binding site (TFBS) to screen the homologous regions ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp077

    authors: Kang K,Chung JH,Kim J

    更新日期:2009-04-01 00:00:00

  • Evolution of a highly functional circular DNA aptamer in serum.

    abstract::Circular DNA aptamers are powerful candidates for therapeutic applications given their dramatically enhanced biostability. Herein we report the first effort to evolve circular DNA aptamers that bind a human protein directly in serum, a complex biofluid. Targeting human thrombin, this strategy has led to the discovery ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa800

    authors: Mao Y,Gu J,Chang D,Wang L,Yao L,Ma Q,Luo Z,Qu H,Li Y,Zheng L

    更新日期:2020-11-04 00:00:00

  • RPI-Pred: predicting ncRNA-protein interaction using sequence and structural information.

    abstract::RNA-protein complexes are essential in mediating important fundamental cellular processes, such as transport and localization. In particular, ncRNA-protein interactions play an important role in post-transcriptional gene regulation like mRNA localization, mRNA stabilization, poly-adenylation, splicing and translation....

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv020

    authors: Suresh V,Liu L,Adjeroh D,Zhou X

    更新日期:2015-02-18 00:00:00