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https://www.readbyqxmd.com/read/27915180/self-primed-isothermal-amplification-for-genomic-dna-detection-of-human-papillomavirus
#1
Wei Lu, Qingpan Yuan, Zhiliu Yang, Bo Yao
Rolling circle amplification (RCA) is an isothermal amplification technique with high efficiency and perfect accuracy for nucleic acids detection. However, RCA technique suffers the limitation to detect short DNA or RNA molecules. For long nucleic acid molecules, enzymatic restriction as well as heat denaturation process is usually required, which makes the amplification not effective and strictly isothermal. In this article, a simple and efficient one-pot self-primed isothermal amplification (SIA) was developed for detection of genomic DNA directly based on the combination of nicking endonuclease assisted strand displacement amplification (SDA) and exponential RCA...
October 10, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27910923/protein-expression-from-unintegrated-hiv-1-dna-introduces-bias-in-primary-in-vitro-post-integration-latency-models
#2
Pawel Bonczkowski, Marie-Angélique De Scheerder, Eva Malatinkova, Alexandra Borch, Zora Melkova, Renate Koenig, Ward De Spiegelaere, Linos Vandekerckhove
To understand the persistence of latently HIV-1 infected cells in virally suppressed infected patients, a number of in vitro models of HIV latency have been developed. In an attempt to mimic the in vivo situation as closely as possible, several models use primary cells and replication-competent viruses in combination with antiretroviral compounds to prevent ongoing replication. Latency is subsequently measured by HIV RNA and/or protein production after cellular activation. To discriminate between pre- and post-integration latency, integrase inhibitors are routinely used, preventing novel integrations upon cellular activation...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27899638/structure-and-folding-of-the-tetrahymena-telomerase-rna-pseudoknot
#3
Darian D Cash, Juli Feigon
Telomerase maintains telomere length at the ends of linear chromosomes using an integral telomerase RNA (TER) and telomerase reverse transcriptase (TERT). An essential part of TER is the template/pseudoknot domain (t/PK) which includes the template, for adding telomeric repeats, template boundary element (TBE), and pseudoknot, enclosed in a circle by stem 1. The Tetrahymena telomerase holoenzyme catalytic core (p65-TER-TERT) was recently modeled in our 9 Å resolution cryo-electron microscopy map by fitting protein and TER domains, including a solution NMR structure of the Tetrahymena pseudoknot...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27883258/bivalent-display-of-dicysteine-on-pna-for-homogenous-dna-rna-detection-via-in-situ-fluorescence-labelling
#4
Gemin Fang, Oliver Seitz
Fluorogenic probes that signal the presence of specific DNA or RNA sequences are key enabling tools for molecular disease diagnosis and imaging studies. Usually, at least one fluorophore is attached via covalent bonds to an oligonucleotide probe. However, the additional conjugation step increases costs. Herein we introduce a method that avoids the requirement for the preparation of fluorescence labeled oligonucleotides and provides the opportunity to alter the fluorogenic reporter dye without resynthesis. The method is based on adjacent hybridization of two dicysteine-containing peptide nucleic acid probes to form a bipartite tetracysteine motif which binds profluorescent bisarsenical dyes such as FIAsH, ReAsH or CrAsH...
November 24, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27822856/in-situ-single-molecule-rna-genotyping-using-padlock-probes-and-rolling-circle-amplification
#5
Tomasz Krzywkowski, Thomas Hauling, Mats Nilsson
Present-day techniques allow for massively parallel and high-throughput characterization of the somatic mutation status of samples. Most of these assays rely on whole specimen extracts, where heterogeneous spatial context of the specimen is lost. This chapter describes an up-to-date protocol for multiplexed, in situ genotyping of RNA in preserved tissue and cell lines, using padlock probes and rolling circle amplification. The presented approach allows for automated quantification of mRNA expression and mutation status, in single cells or in designated specimen areas...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27809244/accumulation-of-stable-full-length-circular-group-i-intron-rnas-during-heat-shock
#6
Kasper L Andersen, Bertrand Beckert, Benoit Masquida, Steinar D Johansen, Henrik Nielsen
Group I introns in nuclear ribosomal RNA of eukaryotic microorganisms are processed by splicing or circularization. The latter results in formation of full-length circular introns without ligation of the exons and has been proposed to be active in intron mobility. We applied qRT-PCR to estimate the copy number of circular intron RNA from the myxomycete Didymium iridis. In exponentially growing amoebae, the circular introns are nuclear and found in 70 copies per cell. During heat-shock, the circular form is up-regulated to more than 500 copies per cell...
October 31, 2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/27797171/the-discovery-of-rolling-circle-amplification-and-rolling-circle-transcription
#7
Michael G Mohsen, Eric T Kool
Nucleic acid amplification is a hugely important technology for biology and medicine. While the polymerase chain reaction (PCR) has been highly useful and effective, its reliance on heating and cooling cycles places some constraints on its utility. For example, the heating step of PCR can destroy biological molecules under investigation and heat/cool cycles are not applicable in living systems. Thus, isothermal approaches to DNA and RNA amplification are under widespread study. Perhaps the simplest of these are the rolling circle approaches, including rolling circle amplification (RCA) and rolling circle transcription (RCT)...
October 24, 2016: Accounts of Chemical Research
https://www.readbyqxmd.com/read/27761361/paradoxical-delay-of-senescence-upon-depletion-of-brca2-in-telomerase-deficient-worms
#8
Mi-Sun Kwon, Jaewon Min, Hee-Yeon Jeon, Kwangwoo Hwang, Chuna Kim, Junho Lee, Je-Gun Joung, Woong-Yang Park, Hyunsook Lee
BRCA2 is a multifunctional tumor suppressor involved in homologous recombination (HR), mitotic checkpoint regulation, and telomere homeostasis. Absence of Brca2 in mice results in progressive shortening of telomeres and senescence, yet cells are prone to neoplastic transformation with elongated telomeres, suggesting that BRCA2 has positive and negative effects on telomere length regulation along the path to tumorigenesis. Using Caenorhabditis elegans as a model, we show here that depletion of BRC-2, an ortholog of BRCA2, paradoxically delays senescence in telomerase-deficient mutant worms...
October 2016: FEBS Open Bio
https://www.readbyqxmd.com/read/27737936/titin-forms-circles-regulation-by-heart-failure-and-the-rna-binding-protein-rbm20
#9
EDITORIAL
Nicolas Jaé, Stefanie Dimmeler
No abstract text is available yet for this article.
October 14, 2016: Circulation Research
https://www.readbyqxmd.com/read/27732664/combined-activity-of-dcl2-and-dcl3-is-crucial-in-the-defense-against-potato-spindle-tuber-viroid
#10
Konstantina Katsarou, Eleni Mavrothalassiti, Wannes Dermauw, Thomas Van Leeuwen, Kriton Kalantidis
Viroids are self replicating non-coding RNAs capable of infecting a wide range of plant hosts. They do not encode any proteins, thus the mechanism by which they escape plant defenses remains unclear. RNAi silencing is a major defense mechanism against virus infections, with the four DCL proteins being principal components of the pathway. We have used Nicotiana benthamiana as a model to study Potato spindle tuber viroid infection. This viroid is a member of the Pospiviroidae family and replicates in the nucleus via an asymmetric rolling circle mechanism...
October 2016: PLoS Pathogens
https://www.readbyqxmd.com/read/27713731/genome-based-genetic-tool-development-for-bacillus-methanolicus-theta-and-rolling-circle-replicating-plasmids-for-inducible-gene-expression-and-application-to-methanol-based-cadaverine-production
#11
Marta Irla, Tonje M B Heggeset, Ingemar Nærdal, Lidia Paul, Tone Haugen, Simone B Le, Trygve Brautaset, Volker F Wendisch
Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27691982/epigenetic-regulation-of-cognition-a-circumscribed-review-of-the-field
#12
Elena L Grigorenko, Sergey A Kornilov, Oksana Yu Naumova
The last decade has been marked by an increased interest in relating epigenetic mechanisms to complex human behaviors, although this interest has not been balanced, accentuating various types of affective and primarily ignoring cognitive functioning. Recent animal model data support the view that epigenetic processes play a role in learning and memory consolidation and help transmit acquired memories even across generations. In this review, we provide an overview of various types of epigenetic mechanisms in the brain (DNA methylation, histone modification, and noncoding RNA action) and discuss their impact proximally on gene transcription, protein synthesis, and synaptic plasticity and distally on learning, memory, and other cognitive functions...
October 3, 2016: Development and Psychopathology
https://www.readbyqxmd.com/read/27663500/foldatlas-a-repository-for-genome-wide-rna-structure-probing-data
#13
Matthew Norris, Chun Kit Kwok, Jitender Cheema, Matthew Hartley, Richard J Morris, Sharon Aviran, Yiliang Ding
: Most RNA molecules form internal base pairs, leading to a folded secondary structure. Some of these structures have been demonstrated to be functionally significant. High-throughput RNA structure chemical probing methods generate millions of sequencing reads to provide structural constraints for RNA secondary structure prediction. At present, processed data from these experiments are difficult to access without computational expertise. Here we present FoldAtlas, a web interface for accessing raw and processed structural data across thousands of transcripts...
September 22, 2016: Bioinformatics
https://www.readbyqxmd.com/read/27662876/a-novel-method-to-quantify-rna-protein-interactions-in-situ-using-fmtrip-and-proximity-ligation
#14
C Zurla, J Jung, E L Blanchard, P J Santangelo
RNA binding proteins (RBP) and small RNAs regulate the editing, localization, stabilization, translation, and degradation of ribonucleic acids (RNAs) through their interactions with specific cis-acting elements within target RNAs. Here, we describe a novel method to detect protein-mRNA interactions, which combines FLAG-peptide modified, multiply-labeled tetravalent RNA imaging probes (FMTRIPs) with proximity ligation (PLA), and rolling circle amplification (RCA). This assay detects native RNA in a sequence specific and single RNA sensitive manner, and PLA allows for the quantification and localization of protein-mRNA interactions with single-interaction sensitivity...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27640431/molecular-characterization-of-a-first-begomovirus-associated-with-lentil-lens-culinaris-from-india
#15
K Naimuddin, M Akram, A K Agnihotri
A disease of lentil with symptoms of distortion, mottling and chlorosis in the leaves, shortening of internodes and excessive branching was noticed in lentil at Kanpur, India, during 2012-2014. Results of polymerase chain reaction and reverse transcriptase polymerase chain reaction employed to detect suspected RNA and DNA viruses indicated involvement of a geminivirus, which was further characterized by sequencing of full genome amplified by rolling circle amplification. Analysis of full length DNA-A revealed 96...
2016: Acta Virologica
https://www.readbyqxmd.com/read/27610229/linking-%C3%AE-to-%C3%AE-diverse-and-dynamic-rna-based-mechanisms-to-regulate-gene-expression-by-5-to-3-communication
#16
REVIEW
Megan E Filbin, Jeffrey S Kieft
Communication between the 5' and 3' ends of a eukaryotic messenger RNA (mRNA) or viral genomic RNA is a ubiquitous and important strategy used to regulate gene expression. Although the canonical interaction between initiation factor proteins at the 5' end of an mRNA and proteins bound to the polyadenylate tail at the 3' end is well known, in fact there are many other strategies used in diverse ways. These strategies can involve "non-canonical" proteins, RNA structures, and direct RNA-RNA base-pairing between distal elements to achieve 5'-to-3' communication...
2016: F1000Research
https://www.readbyqxmd.com/read/27604461/dna-methylation-mediated-up-regulation-of-terra-non-coding-rna-is-coincident-with-elongated-telomeres-in-the-human-placenta
#17
Boris Novakovic, Christine E Napier, Regan Vryer, Eva Dimitriadis, Ursula Manuelpillai, Andrew Sharkey, Jeffrey M Craig, Roger R Reddel, Richard Saffery
STUDY QUESTION: What factors regulate elongated telomere length in the human placenta? SUMMARY ANSWER: Hypomethylation of TERRA promoters in the human placenta is associated with high expression TERRA genes, however, no clear mechanistic link between these phenomena and elongated telomere length in the human placenta was found. WHAT IS KNOWN ALREADY: Human placenta tissue and trophoblasts show longer telomere lengths compared to gestational age-matched somatic cells...
September 7, 2016: Molecular Human Reproduction
https://www.readbyqxmd.com/read/27582462/dna-and-rna-topoisomerase-activities-of-top3%C3%AE-are-promoted-by-mediator-protein-tudor-domain-containing-protein-3
#18
Grace Ee-Lu Siaw, I-Fen Liu, Po-Yen Lin, Michael D Been, Tao-Shih Hsieh
Topoisomerase 3β (Top3β) can associate with the mediator protein Tudor domain-containing protein 3 (TDRD3) to participate in two gene expression processes of transcription and translation. Despite the apparent importance of TDRD3 in binding with Top3β and directing it to cellular compartments critical for gene expression, the biochemical mechanism of how TDRD3 can affect the functions of Top3β is not known. We report here sensitive biochemical assays for the activities of Top3β on DNA and RNA substrates in resolving topological entanglements and for the analysis of TDRD3 functions...
September 20, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27571848/circular-rnas-unexpected-outputs-of-many-protein-coding-genes
#19
Jeremy E Wilusz
Pre-mRNAs from thousands of eukaryotic genes can be non-canonically spliced to generate circular RNAs, some of which accumulate to higher levels than their associated linear mRNA. Recent work has revealed widespread mechanisms that dictate whether the spliceosome generates a linear or circular RNA. For most genes, circular RNA biogenesis via backsplicing is far less efficient than canonical splicing, but circular RNAs can accumulate due to their long half-lives. Backsplicing is often initiated when complementary sequences from different introns base pair and bring the intervening splice sites close together...
August 29, 2016: RNA Biology
https://www.readbyqxmd.com/read/27559075/total-hiv-1-dna-a-marker-of-viral-reservoir-dynamics-with-clinical-implications
#20
REVIEW
Véronique Avettand-Fènoël, Laurent Hocqueloux, Jade Ghosn, Antoine Cheret, Pierre Frange, Adeline Melard, Jean-Paul Viard, Christine Rouzioux
HIV-1 DNA persists in infected cells despite combined antiretroviral therapy (cART), forming viral reservoirs. Recent trials of strategies targeting latent HIV reservoirs have rekindled hopes of curing HIV infection, and reliable markers are thus needed to evaluate viral reservoirs. Total HIV DNA quantification is simple, standardized, sensitive, and reproducible. Total HIV DNA load influences the course of the infection and is therefore clinically relevant. In particular, it is predictive of progression to AIDS and death, independently of HIV RNA load and the CD4 cell count...
October 2016: Clinical Microbiology Reviews
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