bluuebunny/crossref_metadata_2025_split huggingface dataset:
CRISPR-Cas12a-assisted PCR tagging of mammalian genes
Authors: Fueller Julia , Herbst Konrad , Meurer Matthias , Gubicza Krisztina , Kurtulmus Bahtiyar , Knopf Julia D. , Kirrmaier Daniel , Buchmuller Benjamin C. , Pereira Gislene , Lemberg Marius K. , Knop Michael 
Published: November 2018

AbstractHere we describe a time-efficient strategy for endogenous C-terminal gene tagging in mammalian tissue culture cells. An online platform is used to design two long gene-specific oligonucleotides for PCR with generic template cassettes to create linear dsDNA donors, termed PCR cassettes. PCR cassettes encode the tag (e.g. GFP), a Cas12a CRISPR RNA for cleavage of the target locus and short homology arms for directed integration via homologous recombination. The integrated tag is coupled to a generic terminator shielding the tagged gene from the co-inserted auxiliary sequences. Co-transfection of PCR cassettes with a Cas12a-encoding plasmid leads to robust endogenous expression of tagged genes, with tagging efficiency of up to 20% without selection, and up to 60% when selection markers are used. We used target-enrichment sequencing to investigate all potential sources of artefacts. Our work outlines a quick strategy particularly suitable for exploratory studies using endogenous expression of fluorescent protein tagged genes
Neural Signatures of Prediction Errors in a Decision-Making Task Are Modulated by Action Execution Failures
Authors: McDougle Samuel D. , Butcher Peter A. , Parvin Darius , Mushtaq Fasial , Niv Yael , Ivry Richard B. , Taylor Jordan A. 
Published: November 2018

AbstractDecisions must be implemented through actions, and actions are prone to error. As such, when an expected outcome is not obtained, an individual should not only be sensitive to whether the choice itself was suboptimal, but also whether the action required to indicate that choice was executed successfully. The intelligent assignment of credit to action execution versus action selection has clear ecological utility for the learner. To explore this scenario, we used a modified version of a classic reinforcement learning task in which feedback indicated if negative prediction errors were, or were not, associated with execution errors. Using fMRI, we asked if prediction error computations in the human striatum, a key substrate in reinforcement learning and decision making, are modulated when a failure in action execution results in the negative outcome. Participants were more tolerant of non-rewarded outcomes when these resulted from execution errors versus when execution was successful but the reward was withheld. Consistent with this behavior, a model-driven analysis of neural activity revealed an attenuation of the signal associated with negative reward prediction error in the striatum following execution failures. These results converge with other lines of evidence suggesting that prediction errors in the mesostriatal dopamine system integrate high-level information during the evaluation of instantaneous reward outcomes.
Identification of a putative nuclear localization signal in maspin protein shed light into its nuclear import regulation
Authors: Reina Jeffrey , Zhou Lixin , Fontes Marcos R.M. , Panté Nelly , Cella Nathalie 
Published: November 2018

AbstractMaspin (SERPINB5) is a potential tumor suppressor gene with pleiotropic biological activities, including regulation of cell proliferation, death, adhesion, migration and gene expression. Several studies suggest that subcellular localization plays an essential role on maspin tumor suppression activity. In this study we investigated the molecular mechanisms underlying maspin nucleocytoplasmic shuttling. Anin vitronuclear-import assay using digitonin-permeabilized HeLa cells demonstrated that maspin enters the nucleus by an energy-and carrier-independent mechanism. However, previous studies indicated that maspin subcellular localization is regulated in the cell. Using a nuclear localization signal (NLS) prediction software, we identified a putative NLS in the maspin amino acid sequence. To distinguish between passive and regulated nuclear translocation, maspinNLS or the full-length protein (MaspinFL) were fused to 5GFP, rendering the construct too large to enter the nucleus passively. Unexpectedly, 5GFP-maspinNLS, but not maspinFL-5GFP, entered the nucleus of HeLa cells. Dominant-negative Ran-GTPase mutants RanQ69L or RanT24N, suppressed 5GFP-maspinNLS nuclear localization. In summary, we provide evidence that maspin translocates to the nucleus passively. In addition, we identified a peptide in the maspin protein sequence, which is able to drive a 5GFP construct to the nucleus in an energy-dependent manner.
GimmeMotifs: an analysis framework for transcription factor motif analysis
Authors: Bruse Niklas , Heeringen Simon J. van 
Published: November 2018

AbstractBackgroundTranscription factors (TFs) bind to specific DNA sequences, TF motifs, in cis-regulatory sequences and control the expression of the diverse transcriptional programs encoded in the genome. The concerted action of TFs within the chromatin context enables precise temporal and spatial expression patterns. To understand how TFs control gene expression it is essential to model TF binding. TF motif information can help to interpret the exact role of individual regulatory elements, for instance to predict the functional impact of non-coding variants.FindingsHere we present GimmeMotifs, a comprehensive computational framework for TF motif analysis. Compared to the previously published version, this release adds a whole range of new functionality and analysis methods. It now includes tools for de novo motif discovery, motif scanning and sequence analysis, motif clustering, calculation of performance metrics and visualization. Included with GimmeMotifs is a non-redundant database of clustered motifs. Compared to other motif databases, this collection of motifs shows competitive performance in discriminating bound from unbound sequences. Using our de novo motif discovery pipeline we find large differences in performance between de novo motif finders on ChIP-seq data. Using an ensemble method such as implemented in GimmeMotifs will generally result in improved motif identification compared to a single motif finder. Finally, we demonstrate maelstrom, a new ensemble method that enables comparative analysis of TF motifs between multiple high-throughput sequencing experiments, such as ChIP-seq or ATAC-seq. Using a collection of ~200 H3K27ac ChIP-seq data sets we identify TFs that play a role in hematopoietic differentiation and lineage commitment.ConclusionGimmeMotifs is a fully-featured and flexible framework for TF motif analysis. It contains both command-line tools as well as a Python API and is freely available at: https://github.com/vanheeringen-lab/gimmemotifs.
Modulation of RNA-dependent interactions in stress granules prevents persistent TDP-43 accumulation in ALS/FTD
Authors: Fang Mark Y. , Markmiller Sebastian , Dowdle William E. , Vu Anthony Q. , Bushway Paul J. , Ding Sheng , Mercola Mark M. , Lewcock Joseph W. , Yeo Gene W. 
Published: November 2018

ABSTRACTHuman genetic variants are usually represented by four values with variable length: chromosome, position, reference and alternate alleles. Thereis no guarantee that these components are represented in a consistent way across different data sources, and processing variant-based data can be inefficient because four different comparison operations are needed for each variant, three of which are string comparisons. Working with strings, in contrast to numbers, poses extra challenges on computer memory allocation and data-representation. Existing variant identifiers do not typicallyrepresent every possible variant we may be interested in, nor they are directly reversible. To overcome these limitations,VariantKey, a novel reversible numerical encoding schema for human genetic variants, is presented here alongside a multi-language open-source software implementation (http://github.com/genomicspls/variantkey). VariantKey represents variants as single 64 bit numeric entities, while preserving the ability to be searched and sorted by chromosome and position. The individual components of short variants can be directly read back from the VariantKey, while long variants are supported with a fast lookup table.Highlights~100 compounds identified by high-content screen inhibit SGs in HEK293, NPCs and iPS-MNs.ALS-associated RBPs are recruited to SGs in an RNA-dependent mannerMolecules with planar moieties prevent recruitment of ALS-associated RBPs to SGsCompounds inhibit TDP-43 accumulation in SGs and inTARDBPmutant iPS-MNs.
Annotations capturing cell-type-specific TF binding explain a large fraction of disease heritability
Authors: Geijn Bryce van de , Finucane Hilary , Gazal Steven , Hormozdiari Farhad , Amariuta Tiffany , Liu Xuanyao , Gusev Alexander , Loh Po-Ru , Reshef Yakir , Kichaev Gleb , Raychauduri Soumya , Price Alkes L. 
Published: November 2018

AbstractIt is widely known that regulatory variation plays a major role in complex disease and that cell-type-specific binding of transcription factors (TF) is critical to gene regulation, but genomic annotations from directly measured TF binding information are not currently available for most cell-type-TF pairs. Here, we construct cell-type-specific TF binding annotations by intersecting sequence-based TF binding predictions with cell-type-specific chromatin data; this strategy addresses both the limitation that identical sequences may be bound or unbound depending on surrounding chromatin context, and the limitation that sequence-based predictions are generally not cell-type-specific. We evaluated different combinations of sequence-based TF predictions and chromatin data by partitioning the heritability of 49 diseases and complex traits (average N=320K) using stratified LD score regression with the baseline-LD model (which is not cell-type-specific). We determined that 100bp windows around MotifMap sequenced-based TF binding predictions intersected with a union of six cell-type-specific chromatin marks (imputed using ChromImpute) performed best, with an 58% increase in heritability enrichment compared to the chromatin marks alone (11.6x vs 7.3x; P = 9 × 10-14 for difference) and a 12% increase in cell-type-specific signal conditional on annotations from the baseline-LD model (P = 8 × 10-11 for difference). Our results show that intersecting sequence-based TF predictions with cell-type-specific chromatin information can help refine genome-wide association signals.
Artificial selection of microbial communities to enhance degradation of recalcitrant polymers
Authors: Wright Robyn J. , Gibson Matthew I. , Christie-Oleza Joseph A. 
Published: November 2018

AbstractRecalcitrant polymers are widely distributed in the environment. This includes natural polymers, such as chitin, but synthetic polymers are becoming increasingly abundant, for which biodegradation is uncertain. Distribution of labour in microbial communities commonly evolves in nature, particularly for arduous processes, suggesting a community may be better at degrading recalcitrant compounds than individual microorganisms. Artificial selection of microbial communities with better degradation potential has seduced scientists for over a decade, but the method has not been systematically optimised nor applied to polymer degradation. Using chitin as a case study, we successfully selected for microbial communities with enhanced chitinase activities but found that continuous optimisation of incubation times between selective generations was of utmost importance. The analysis of the community composition over the entire selection process revealed fundamental aspects in microbial ecology: when incubation times between generations were optimal, the system was dominated byGammaproteobacteria, main bearers of chitinase enzymes and drivers of chitin degradation, before being succeeded by cheating, cross-feeding and grazing organisms.ImportanceArtificial selection is a powerful and atractive technique that can enhance the biodegradation of a recalcitrant polymer and other pollutants by microbial communities. We show, for the first time, that the success of artificially selecting microbial communities requires an optimisation of the incubation times between generations when implementing this method. Hence, communities need to be transferred at the peak of the desired activity in order to avoid community drift and replacement of the efficient biodegrading community by cheaters, cross-feeders and grazers.
Re-expression of SynGAP Protein in Adulthood Improves Translatable Measures of Brain Function and Behavior in a Model of Neurodevelopmental Disorders
Authors: Creson Thomas K. , Rojas Camilo , Hwaun Ernie , Vaissiere Thomas , Kilinc Murat , Holder J. Lloyd , Tang Jianrong , Colgin Laura Lee , Miller Courtney A. , Rumbaugh Gavin 
Published: November 2018

AbstractBackgroundNeurodevelopmental disorder (NDD) risk genes have pleiotropic biological functions, such as control over both developmental and non-developmental processes that influence disease-related phenotypes. Currently, it remains unclear how developmental versus non-developmental processes influence the duration and/or effectiveness of permissive treatment windows for NDDs. SYNGAP1 haploinsufficiency causes an NDD defined by autistic traits, cognitive impairment, and epilepsy. Syngap1 heterozygosity in mice disrupts a developmental critical period, and, consistent with this, certain behavioral abnormalities are resistant to gene therapy initiated in adulthood. However, the Syngap1 endophenotype is extensive and this protein has diverse cell biological functions. Therefore, SynGAP pleiotropy may influence the permissive treatment window for previously untested disease-relevant phenotypes.MethodsA whole-body gene restoration technique was used to determine how restoration of SynGAP protein in adult heterozygous mice impacted previously untested phenotypes, such as memory, seizure susceptibility, systems-level cortical hyperexcitability, and hippocampal oscillations linked to mnemonic processes.ResultsAdult restoration of SynGAP protein in haploinsufficient mice reversed long-term contextual memory deficits and behavioral measures of seizure susceptibility. Moreover, SynGAP re-expression in adult mice eliminated brain state-dependent, patient-linked paroxysmal interictal spiking and increased the amplitude of hippocampal theta oscillations.ConclusionsSynGAP protein in the mature brain dynamically regulates neural circuit function and influences disease-relevant phenotypes. The impact of these findings is that treatments targeting certain debilitating aspects of SYNGAP1-related disorders may be effective throughout life. Moreover, the efficacy of experimental treatments for SYNGAP1 patients may be quantifiable through changes in species-conserved, state-dependent pathological electroencephalogram signals.
CRISPR-TSKO facilitates efficient cell type-, tissue-, or organ-specific mutagenesis in Arabidopsis
Authors: Decaestecker Ward , Buono Rafael Andrade , Pfeiffer Marie L. , Vangheluwe Nick , Jourquin Joris , Karimi Mansour , Van Isterdael Gert , Beeckman Tom , Nowack Moritz K. , Jacobs Thomas B. 
Published: November 2018

AbstractDetailed functional analyses of many fundamentally-important plant genes via conventional loss-of-function approaches are impeded by severe pleiotropic phenotypes. In particular, mutations in genes that are required for basic cellular functions and/or reproduction often interfere with the generation of homozygous mutant plants, precluding further functional studies. To overcome this limitation, we devised a CRISPR-based tissue-specific knockout system, CRISPR-TSKO, enabling the generation of somatic mutations in particular plant cell types, tissues, and organs. In Arabidopsis, CRISPR-TSKO mutations in essential genes caused well-defined, localized phenotypes in the root cap, stomatal lineage, or entire lateral roots. The underlying modular cloning system allows for efficient selection, identification, and functional analysis of mutant lines directly in the first transgenic generation. The efficacy of CRISPR-TSKO opens new avenues to discover and analyze gene functions in spatial and temporal contexts of plant life while avoiding pleiotropic effects of system-wide loss of gene function.
Intrinsically disordered regions regulate both catalytic and non-catalytic activities of the MutLα mismatch repair complex
Authors: Kim Yoori , Furman Christopher M. , Manhart Carol M. , Alani Eric , Finkelstein Ilya J. 
Published: November 2018

AbstractIntrinsically disordered regions (IDRs) are present in at least 30% of the eukaryotic proteome and are enriched in chromatin-associated proteins. Using a combination of genetics, biochemistry, and single-molecule biophysics, we characterize how IDRs regulate the functions of the yeast MutLα (Mlh1-Pms1) mismatch repair (MMR) complex. Shortening or scrambling the IDRs in both subunits ablates MMR in vivo. Mlh1-Pms1 complexes with shorter IDRs that disrupt MMR retain wild-type DNA binding affinity but are impaired for diffusion on both naked and nucleosome-coated DNA. Moreover, the IDRs also regulate the ATP hydrolysis and nuclease activities that are encoded in the structured N- and C-terminal domains of the complex. This combination of phenotypes underlies the catastrophic MMR defect seen with the mutant MutLα in vivo. More broadly, this work highlights an unanticipated multi-functional role for IDRs in regulating both facilitated diffusion on chromatin and nucleolytic processing of a DNA substrate.
Metagenomic Insights Unveil the Dominance of Undescribed Actinobacteria in Pond Ecosystem of an Indian Shrine
Authors: Manikkam Radhakrishnan , Imchen Madangchanok , Kaari Manigundan , Vignesh Angamuthu , Gopikrishnan Venugopal , Shanmugasundaram Thangavel , Joseph Jerrine , Balagurunathan Ramasamy , Kumavath Ranjith 
Published: November 2018

AbstractMetagenomic analysis holds immense potential for identifying rare and uncharacterized microorganisms from many ecological habitats. Actinobacteria have been proved to be an excellent source of novel antibiotics for several decades. The present study was designed to delineate and understand the bacterial diversity with special focus on Actinobacteria from pond sediment collected from Sanjeeviraya Hanuman Temple, Ayyangarkulam, Kanchipuram, Tamil Nadu, India. The sediment had an average temperature (25.32%), pH (7.13), salinity (0.960 mmhos/cm) and high organic content (10.7%) posing minimal stress on growth condition of the microbial community. Subsequent molecular manipulations, sequencing and bioinformatics analysis of V3 and V4 region of 16S rRNA metagenomics analysis confirmed the presence of 40 phyla, 100 classes, 223 orders, 319 families and 308 genera in the sediment sample dominated by Acidobacteria (18.14%), Proteobacteria (15.13%), Chloroflexi (12.34), Actinobacteria (10.84%), Cyanobacteria (5.58%), Verrucomicrobia (3.37%), Firmicutes (2.28%), and, Gemmatimonadetes (1.63%). Among the Actinobacteria phylum, Acidothermus (29.68%) was the predominant genus followed by Actinospica (17.65%), Streptomyces (14.64%), Nocardia (4.55%) and Sinomonas (2.9%). Culture-dependent isolation of Actinobacteria yielded all strains of similar morphology to that of Streptomyces genus which clearly indicating that the traditional based technique is incapable of isolating majority of the non-Streptomyces or the so called rare Actinobacteria. Although Actinobacteria were among the dominant phylum, a close look at the species level indicated that only 15.2% within the Actinobacterial phylum could be assigned to cultured species. This leaves a vast majority of the Actinobacterial species yet to be explored with possible novel metabolites have special pharmaceutical and industrial application. It also indicates that the microbial ecology of pond sediment is neglected fields which need attention.
Spatio-temporal dynamics of dengue in Brazil: seasonal travelling waves and determinants of regional synchrony
Authors: Churakov Mikhail , Villabona-Arenas Christian J. , Kraemer Moritz U.G. , Salje Henrik , Cauchemez Simon 
Published: November 2018

AbstractDengue continues to be the most important vector-borne viral disease globally and in Brazil, where more than 1.4 million cases and over 500 deaths were reported in 2016. Mosquito control programmes and other interventions have not stopped the alarming trend of increasingly large epidemics in the past few years.Here, we analyzed monthly dengue cases reported in Brazil between 2001 and 2016 to better characterize the key drivers of dengue epidemics. Spatio-temporal analysis revealed recurring travelling waves of disease occurrence. Using wavelet methods, we characterised the average seasonal pattern of dengue in Brazil, which starts in the western states of Acre and Rondônia, then travels eastward to the coast before reaching the northeast of the country. Only two states in the north of Brazil (Roraima and Amapá) did not follow the countrywide pattern and had inconsistent timing of dengue epidemics throughout the study period.We also explored epidemic synchrony and timing of annual dengue cycles in Brazilian regions. Using gravity style models combined with climate factors, we showed that both human mobility and vector ecology contribute to spatial patterns of dengue occurrence.This study offers a characterization of the spatial dynamics of dengue in Brazil and its drivers, which could inform intervention strategies against dengue and other arboviruses.Author summaryIn this paper we studied the synchronization of dengue epidemics in Brazilian regions. We found that a typical dengue season in Brazil can be described as a wave travelling from the western part of the country towards the east, with the exception of the two most northern equatorial states that experienced inconsistent seasonality of dengue epidemics.We found that the spatial structure of dengue cases is driven by both climate and human mobility patterns. In particular, precipitation was the most important factor for the seasonality of dengue at finer spatial resolutions.Our findings increase our understanding of large scale dengue patterns and could be used to enhance national control programs against dengue and other arboviruses.