Light-activated RNA interference using double-stranded siRNA precursors modified using a remarkable regiospecificity of diazo-based photolabile groups.

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 the phosphate backbone of RNA and DNA. In this work we find that these diazo-based reagents in fact react very poorly with backbone phosphates. Instead, they show a remarkable specificity for terminal phosphates and very modest modification of the nucleobases. Furthermore, the photo deprotection of these terminal modifications is shown to be much more facile than nucleobase modified sites. In this study we have characterized this regiospecificity using RNA duplexes and model nucleotides, analyzed using LC/MS/MS. We have also applied this understanding of the regio-specificity to our technique of light activated RNA interference (LARI). We examined 27-mer double-stranded precursors of siRNA ('dsRNA'), and have modified them using the photo-cleavable di-methoxy nitro phenyl ethyl group (DMNPE) group. By incorporating terminal phosphates in the dsRNA, we are able to guide DMNPE to react at these terminal locations. These modified dsRNA duplexes show superior performance to our previously described DMNPE-modified siRNA, with the range of expression that can be toggled by light increasing by a factor of two.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Shah S,Jain PK,Kala A,Karunakaran D,Friedman SH

doi

10.1093/nar/gkp415

subject

Has Abstract

pub_date

2009-07-01 00:00:00

pages

4508-17

issue

13

eissn

0305-1048

issn

1362-4962

pii

gkp415

journal_volume

37

pub_type

杂志文章
  • Expression of herpes virus thymidine kinase in Neurospora crassa.

    abstract::The expression of thymidine kinase in fungi, which normally lack this enzyme, will greatly aid the study of DNA metabolism and provide useful drug-sensitive phenotypes. The herpes simplex virus type-1 thymidine kinase gene ( tk ) was expressed in Neurospora crassa. tk was expressed as a fusion to N.crassa arg-2 regula...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/25.12.2389

    authors: Sachs MS,Selker EU,Lin B,Roberts CJ,Luo Z,Vaught-Alexander D,Margolin BS

    更新日期:1997-06-15 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

  • MRPrimer: a MapReduce-based method for the thorough design of valid and ranked primers for PCR.

    abstract::Primer design is a fundamental technique that is widely used for polymerase chain reaction (PCR). Although many methods have been proposed for primer design, they require a great deal of manual effort to generate feasible and valid primers, including homology tests on off-target sequences using BLAST-like tools. That ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv632

    authors: Kim H,Kang N,Chon KW,Kim S,Lee N,Koo J,Kim MS

    更新日期:2015-11-16 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

  • pre-miRNA profiles obtained through application of locked nucleic acids and deep sequencing reveals complex 5'/3' arm variation including concomitant cleavage and polyuridylation patterns.

    abstract::Recent research hints at an underappreciated complexity in pre-miRNA processing and regulation. Global profiling of pre-miRNA and its potential to increase understanding of the pre-miRNA landscape is impeded by overlap with highly expressed classes of other non coding (nc) RNA. Here, we present a data set excluding th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr903

    authors: Burroughs AM,Kawano M,Ando Y,Daub CO,Hayashizaki Y

    更新日期:2012-02-01 00:00:00

  • A cellular protein binds to a conserved sequence in the adenovirus type 2 enhancer.

    abstract::A sensitive gel retention assay has been utilized to detect proteins from uninfected Hela nuclei which interact with the adenovirus type 2 enhancer. This assay has been employed to monitor fractionation of nuclear extracts. Three enhancer binding factors were resolved by chromatography on DEAE-Sepharose and one of the...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/15.6.2719

    authors: Barrett P,Clark L,Hay RT

    更新日期:1987-03-25 00:00:00

  • Molecular structures of two new anti-HIV nucleoside analogs: 9-(2,3-dideoxy-2-fluoro-beta-D-threo-pentofuranosyl)adenine and 9-(2,3-dideoxy-2-fluoro-beta-D-threo-pentofuranosyl)hypoxanthine.

    abstract::The x-ray crystal structures of two new anti-HIV compounds, 9-(2,3-dideoxy-2-fluoro-beta-D-threo-pentofuranosyl)adenine (2'-F-dd-araA) and 9-(2,3-dideoxy-2-fluoro-beta-D-threo- pentofuranosyl)hypoxanthine (2'-F-dd-aral), have been determined at two temperatures. Both crystals are in the space group P2(1)2(1)2(1), and ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/20.3.459

    authors: Liaw YC,Gao YG,Marquez VE,Wang AH

    更新日期:1992-02-11 00:00:00

  • B lymphocyte-specific, Cre-mediated mutagenesis in mice.

    abstract::Adaptation of the P1 phage-derived Cre /loxP site- specific recombination system to the gene targeting technique allows for the conditional deletion of genes in mice. To selectively modify genes in B lymphocytes, we have generated mice (designated CD19-Cre) which express cre under the transcriptional control of the B ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/25.6.1317

    authors: Rickert RC,Roes J,Rajewsky K

    更新日期:1997-03-15 00:00:00

  • Flexible expressed region analysis for RNA-seq with derfinder.

    abstract::Differential expression analysis of RNA sequencing (RNA-seq) data typically relies on reconstructing transcripts or counting reads that overlap known gene structures. We previously introduced an intermediate statistical approach called differentially expressed region (DER) finder that seeks to identify contiguous regi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw852

    authors: Collado-Torres L,Nellore A,Frazee AC,Wilks C,Love MI,Langmead B,Irizarry RA,Leek JT,Jaffe AE

    更新日期:2017-01-25 00:00:00

  • Investigating the role of the Est3 protein in yeast telomere replication.

    abstract::The Est3 subunit of yeast telomerase, which adopts a predicted OB-fold, is essential for telomere replication. To assess the possible contributions that Est3 might make to enzyme catalysis, we compared telomerase activity from wild type and est3-Delta strains of Saccharomyces castellii, which revealed that loss of the...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp1173

    authors: Lee J,Mandell EK,Rao T,Wuttke DS,Lundblad V

    更新日期:2010-04-01 00:00:00

  • Nucleotide sequence of the EcoRI D fragment of adenovirus 2 genome.

    abstract::The entire nucleotide sequence of the Ad. 2 EcoRI D fragment has been determined using the Maxam and Gilbert method. This sequence of 2678 bp contains informations relative to late mRNAs ending at position 78 and for which an AATAAA sequence corresponding to their 3' ends is found at residue number 833. Position of th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/8.10.2173

    authors: Hérissé J,Courtois G,Galibert F

    更新日期:1980-05-24 00:00:00

  • Promoter and regulon analysis of nitrogen assimilation factor, sigma54, reveal alternative strategy for E. coli MG1655 flagellar biosynthesis.

    abstract::Bacteria core RNA polymerase (RNAP) must associate with a sigma factor to recognize promoter sequences. Promoters recognized by the sigma(54) (or sigma(N)) associated RNA polymerase are unique in having conserved positions around -24 and -12 nucleotides upstream from the transcriptional start site. Using DNA microarra...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp1123

    authors: Zhao K,Liu M,Burgess RR

    更新日期:2010-03-01 00:00:00

  • Cloning and sequencing of a full length cDNA coding for human retinol-binding protein.

    abstract::We have isolated and sequenced a cDNA clone coding for human Retinol Binding Protein. The sequence indicates that Retinol Binding Protein is synthesized as a single polypeptide chain precursor which is then matured to the secreted protein by removal of a leader peptide. Southern and Northern blot analysis suggest that...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/11.22.7769

    authors: Colantuoni V,Romano V,Bensi G,Santoro C,Costanzo F,Raugei G,Cortese R

    更新日期:1983-11-25 00:00:00

  • Yeast two-hybrid junk sequences contain selected linear motifs.

    abstract::Yeast two-hybrid (Y2H) screenings result in identification of many out-of-frame (OOF) clones that code for short (2-100 amino acids) peptides with no sequence homology to known proteins. We hypothesize that these peptides can reveal common short linear motifs (SLiMs) responsible for their selection. We present a new p...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr600

    authors: Liu Y,Woods NT,Kim D,Sweet M,Monteiro AN,Karchin R

    更新日期:2011-10-01 00:00:00

  • G-quadruplexes and G-quadruplex ligands: targets and tools in antiviral therapy.

    abstract::G-quadruplexes (G4s) are non-canonical nucleic acids secondary structures that form within guanine-rich strands of regulatory genomic regions. G4s have been extensively described in the human genome, especially in telomeres and oncogene promoters; in recent years the presence of G4s in viruses has attracted increasing...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,评审

    doi:10.1093/nar/gky187

    authors: Ruggiero E,Richter SN

    更新日期:2018-04-20 00:00:00

  • Sequence data handling by computer.

    abstract::The speed of the new DNA sequencing techniques has created a need for computer programs to handle the data produced. This paper describes simple programs designed specifically for use by people with little or no computer experience. The programs are for use on small computers and provide facilities for storage, editin...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/4.11.4037

    authors: Staden R

    更新日期:1977-11-01 00:00:00

  • Protein p56 from the Bacillus subtilis phage phi29 inhibits DNA-binding ability of uracil-DNA glycosylase.

    abstract::Protein p56 (56 amino acids) from the Bacillus subtilis phage 29 inactivates the host uracil-DNA glycosylase (UDG), an enzyme involved in the base excision repair pathway. At present, p56 is the only known example of a UDG inhibitor encoded by a non-uracil containing viral DNA. Using analytical ultracentrifugation met...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm584

    authors: Serrano-Heras G,Ruiz-Masó JA,del Solar G,Espinosa M,Bravo A,Salas M

    更新日期:2007-01-01 00:00:00

  • A bichromatic fluorescent reporter for cell-based screens of alternative splicing.

    abstract::Alternative splicing is the primary source of proteome complexity in metazoans and its regulation shapes the proteome in response to shifting physiological requirements. We developed a bichromatic splicing reporter that uses a peculiar feature of some fluorescent protein coding regions to express two different fluores...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl967

    authors: Orengo JP,Bundman D,Cooper TA

    更新日期:2006-01-01 00:00:00

  • A deafness-associated tRNA mutation caused pleiotropic effects on the m1G37 modification, processing, stability and aminoacylation of tRNAIle and mitochondrial translation.

    abstract::Defects in the posttranscriptional modifications of mitochondrial tRNAs have been linked to human diseases, but their pathophysiology remains elusive. In this report, we investigated the molecular mechanism underlying a deafness-associated tRNAIle 4295A>G mutation affecting a highly conserved adenosine at position 37,...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa1225

    authors: Meng F,Zhou M,Xiao Y,Mao X,Zheng J,Lin J,Lin T,Ye Z,Cang X,Fu Y,Wang M,Guan MX

    更新日期:2021-01-25 00:00:00

  • Comprehensive mapping of the human cytokine gene regulatory network.

    abstract::Proper cytokine gene expression is essential in development, homeostasis and immune responses. Studies on the transcriptional control of cytokine genes have mostly focused on highly researched transcription factors (TFs) and cytokines, resulting in an incomplete portrait of cytokine gene regulation. Here, we used enha...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa1055

    authors: Santoso CS,Li Z,Lal S,Yuan S,Gan KA,Agosto LM,Liu X,Pro SC,Sewell JA,Henderson A,Atianand MK,Fuxman Bass JI

    更新日期:2020-12-02 00:00:00

  • Modulation of ATR-mediated DNA damage checkpoint response by cryptochrome 1.

    abstract::Mammalian cryptochromes (Crys) are essential circadian clock factors implicated in diverse clock-independent physiological functions, including DNA damage responses. Here we show that Cry1 modulates the ATR-mediated DNA damage checkpoint (DDC) response by interacting with Timeless (Tim) in a time-of-day-dependent mann...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku094

    authors: Kang TH,Leem SH

    更新日期:2014-04-01 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

  • Intramolecular quadruplex conformation of human telomeric DNA assessed with 125I-radioprobing.

    abstract::A repeated non-coding DNA sequence d(TTAGGG)n is present in the telomeric ends of all human chromosomes. These repeats can adopt multiple inter and intramolecular non-B-DNA conformations that may play an important role in biological processes. Two intramolecular structures of the telomeric oligonucleotide dAGGG(TTAGGG...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkh875

    authors: He Y,Neumann RD,Panyutin IG

    更新日期:2004-10-08 00:00:00

  • Quantitative analysis of condensation/decondensation status of pDNA in the nuclear sub-domains by QD-FRET.

    abstract::Recent studies indicate that controlling the nuclear decondensation and intra-nuclear localization of plasmid DNA (pDNA) would result in an increased transfection efficiency. In the present study, we established a technology for imaging the nuclear condensation/decondensation status of pDNA in nuclear subdomains using...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq1327

    authors: Shaheen SM,Akita H,Yamashita A,Katoono R,Yui N,Biju V,Ishikawa M,Harashima H

    更新日期:2011-04-01 00:00:00

  • Fractionation and characterization of satellite DNAs of the kangaroo rat (Dipodomys ordii).

    abstract::Nuclear DNA from liver cells of the kangaroo rat species Dipodomysordii was fractionated and characterized with the aid of buoyant density gradients in neutral and alkaline CsCl and in Ag(+)-Cs(2)SO(4). More than one-half of the DNA was present in three density satellites, a greater proportion than in any other specie...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/1.4.559

    authors: Hatch FT,Mazrimas JA

    更新日期:1974-04-01 00:00:00

  • 31P-NMR analysis of the B to Z transition in double-stranded (dC-dG)3 and (dC-dG)4 in high salt solution.

    abstract::In 4M NaCl solutions (dC-dG)n (n = 3,4; approximately 9 mM) exist as a mixture o +/- B and Z forms. The low and high field components of two 31P NMR resonances originating from internal phosphodiester groups are assigned to the GpC and CpG linkages, respectively. Low temperatures stabilize the Z-forms, which completel...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.11.4625

    authors: Holak TA,Borer PN,Levy GC,van Boom JH,Wang AH

    更新日期:1984-06-11 00:00:00

  • DNA methylation at a CCGG sequence in the large intron of the rabbit beta-globin gene: tissue-specific variations.

    abstract::We have analysed DNA modification in a HapII site (CCGG) present in the major intron of the discontinuous rabbit beta-globin gene. In most somatic tissues, including erythroid and non-erythroid tissues, about 50% of the DNA is resistant to cleavage at this site by HapII, though 100% cleavage is found with the isoschiz...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/5.12.4631

    authors: Waalwijk C,Flavell RA

    更新日期:1978-12-01 00:00:00

  • Multiple functions for the N-terminal region of Msh6.

    abstract::The eukaryotic mismatch repair protein Msh6 shares five domains in common with other MutS members. However, it also contains several hundred additional residues at its N-terminus. A few of these residues bind to PCNA, but the functions of the other amino acids in the N-terminal region (NTR) are unknown. Here we demons...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm409

    authors: Clark AB,Deterding L,Tomer KB,Kunkel TA

    更新日期:2007-01-01 00:00:00

  • Conserved principles of mammalian transcriptional regulation revealed by RNA half-life.

    abstract::RNA levels in a cell are regulated by the relative rates of RNA synthesis and decay. We recently developed a new approach for measuring both RNA synthesis and decay in a single experimental setting by biosynthetic labeling of newly transcribed RNA. Here, we show that this provides measurements of RNA half-lives from m...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkp542

    authors: Friedel CC,Dölken L,Ruzsics Z,Koszinowski UH,Zimmer R

    更新日期:2009-09-01 00:00:00

  • ATTED-II: a database of co-expressed genes and cis elements for identifying co-regulated gene groups in Arabidopsis.

    abstract::Publicly available database of co-expressed gene sets would be a valuable tool for a wide variety of experimental designs, including targeting of genes for functional identification or for regulatory investigation. Here, we report the construction of an Arabidopsis thaliana trans-factor and cis-element prediction data...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl783

    authors: Obayashi T,Kinoshita K,Nakai K,Shibaoka M,Hayashi S,Saeki M,Shibata D,Saito K,Ohta H

    更新日期:2007-01-01 00:00:00