scholarly journals BarkBase: Epigenomic Annotation of Canine Genomes

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 433 ◽  
Author(s):  
Kate Megquier ◽  
Diane P. Genereux ◽  
Jessica Hekman ◽  
Ross Swofford ◽  
Jason Turner-Maier ◽  
...  
Keyword(s):  
Regulatory Elements ◽  
Open Chromatin ◽  
Genetic Associations ◽  
Functional Studies ◽  
Genomic Annotation ◽  
File Formats ◽  
Causal Variants ◽  
Health And Disease ◽  
Differential Gene ◽  
Accessible Chromatin

Dogs are an unparalleled natural model for investigating the genetics of health and disease, particularly for complex diseases like cancer. Comprehensive genomic annotation of regulatory elements active in healthy canine tissues is crucial both for identifying candidate causal variants and for designing functional studies needed to translate genetic associations into disease insight. Currently, canine geneticists rely primarily on annotations of the human or mouse genome that have been remapped to dog, an approach that misses dog-specific features. Here, we describe BarkBase, a canine epigenomic resource available at barkbase.org. BarkBase hosts data for 27 adult tissue types, with biological replicates, and for one sample of up to five tissues sampled at each of four carefully staged embryonic time points. RNA sequencing is complemented with whole genome sequencing and with assay for transposase-accessible chromatin using sequencing (ATAC-seq), which identifies open chromatin regions. By including replicates, we can more confidently discern tissue-specific transcripts and assess differential gene expression between tissues and timepoints. By offering data in easy-to-use file formats, through a visual browser modeled on similar genomic resources for human, BarkBase introduces a powerful new resource to support comparative studies in dogs and humans.

scholarly journals Chromatin accessibility and regulatory vocabulary across indicine cattle tissues

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Pâmela A. Alexandre ◽  
Marina Naval-Sánchez ◽  
Moira Menzies ◽  
Loan T. Nguyen ◽  
Laercio R. Porto-Neto ◽  
...  
Keyword(s):  
Motif Discovery ◽  
Target Genes ◽  
Developmental Stages ◽  
Bos Taurus ◽  
Regulatory Elements ◽  
Chromatin Accessibility ◽  
Model Organisms ◽  
Open Chromatin ◽  
Health And Disease ◽  
Collaborative Efforts

Abstract Background Spatiotemporal changes in the chromatin accessibility landscape are essential to cell differentiation, development, health, and disease. The quest of identifying regulatory elements in open chromatin regions across different tissues and developmental stages is led by large international collaborative efforts mostly focusing on model organisms, such as ENCODE. Recently, the Functional Annotation of Animal Genomes (FAANG) has been established to unravel the regulatory elements in non-model organisms, including cattle. Now, we can transition from prediction to validation by experimentally identifying the regulatory elements in tropical indicine cattle. The identification of regulatory elements, their annotation and comparison with the taurine counterpart, holds high promise to link regulatory regions to adaptability traits and improve animal productivity and welfare. Results We generate open chromatin profiles for liver, muscle, and hypothalamus of indicine cattle through ATAC-seq. Using robust methods for motif discovery, motif enrichment and transcription factor binding sites, we identify potential master regulators of the epigenomic profile in these three tissues, namely HNF4, MEF2, and SOX factors, respectively. Integration with transcriptomic data allows us to confirm some of their target genes. Finally, by comparing our results with Bos taurus data we identify potential indicine-specific open chromatin regions and overlaps with indicine selective sweeps. Conclusions Our findings provide insights into the identification and analysis of regulatory elements in non-model organisms, the evolution of regulatory elements within two cattle subspecies as well as having an immediate impact on the animal genetics community in particular for a relevant productive species such as tropical cattle.

scholarly journals Disruption of c-MYC binding and chromosomal looping involving genetic variants associated with ankylosing spondylitis upstream of RUNX3 promoter

2021 ◽  
Author(s):  
Carla J Cohen ◽  
Connor Davidson ◽  
Carlo Selmi ◽  
Paul Bowness ◽  
Julian C Knight ◽  
...  
Keyword(s):  
Ankylosing Spondylitis ◽  
Binding Site ◽  
Regulatory Elements ◽  
Published Data ◽  
Open Chromatin ◽  
Physical Interaction ◽  
Nucleotide Polymorphisms ◽  
Genetic Associations ◽  
Chromosome Conformation ◽  
Nuclear Extracts

ABSTRACTBackgroundAnkylosing Spondylitis (AS) is a common form of inflammatory spinal arthritis with a complex aetiology and high heritability, involving more than 100 genetic associations. These include several AS-associated single nucleotide polymorphisms (SNPs) upstream of RUNX3, which encodes the multifunctional RUNT-related transcription factor (TF) 3. The lead associated SNP rs6600247 (p= 2.6 x 10-15) lies ~13kb upstream of the RUNX3 promoter adjacent to a c-MYC TF binding-site. The effect of rs6600247 genotype on DNA binding and chromosome looping were investigated by electrophoretic mobility gel shift assays (EMSA), Western blotting-EMSA (WEMSA) and Chromosome Conformation Capture (3C).ResultsInterrogation of ENCODE published data showed open chromatin in the region overlapping rs6600247 in primary human CD14+ monocytes in contrast to Jurkat T cell line or primary T-cells. The rs6600247 AS-risk allele is predicted to specifically disrupt a c-MYC binding-site. Using a 50bp DNA probe spanning rs6600247 there was consistently less binding to the AS-risk “C” allele of both purified c-MYC protein and nuclear extracts (NE) from monocyte-like U937 cells. WEMSA on U937 NE and purified c-MYC protein confirmed these differences (n=2; p<0.05). 3C experiments demonstrated negligible interaction between the region encompassing rs6600247 and the RUNX3 promoter. A stronger interaction frequency was demonstrated between the RUNX3 promoter and the previously characterised AS-associated SNP rs4648889.ConclusionsThe lead SNP rs6600247, located in an enhancer-like region upstream of the RUNX3 promoter, modulates c-MYC binding. However, the region encompassing rs6600247 has rather limited physical interaction with the promoter of RUNX3. In contrast a clear chromatin looping event between the region encompassing rs4648889 and the RUNX3 promoter was observed. These data provide further evidence for complexity in the regulatory elements upstream of the RUNX3 promoter and the involvement of RUNX3 transcriptional regulation in AS.

scholarly journals HiCoP, a simple and robust method for detecting interactions of regulatory regions

2020 ◽  
Author(s):  
Yan Zhang ◽  
Zhaoqiang Li ◽  
Shasha Bian ◽  
Hao Zhao ◽  
Delong Feng ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Regulatory Elements ◽  
High Signal ◽  
Open Chromatin ◽  
Regulatory Regions ◽  
Essential Information ◽  
Histone H3k27 ◽  
Development And Differentiation ◽  
Physical Interactions ◽  
Accessible Chromatin

Abstract Background: Chromatin physical interactions provides essential information for understanding the regulation of cis -elements like enhancers, promoters, and insulators in cell development and differentiation. The Hi-C assay is a technique detecting chromatin structures of the whole genome but not sensitive to interactions of regulatory elements. Several methods, like HiChIP, DNase-C, and OCEAN-C, have been developed for enriching interactions of regulatory regions, but all of them have some shortcomings. New simple, efficient, and robust methods are still in need of detecting interactions of regulatory regions. Results: We developed a new, simple, and robust assay called CoP ( Co lumn P urified chromatin) for profiling of open chromatin regions by directly purifying fragmentized crosslinked chromatin with a DNA purification column. The open chromatin regions, including active enhancers, promoters, and insulators, were significantly enriched in CoP chromatin. The CoP-seq assay can efficiently detect open chromatin regions, especially active promoters, with a high signal-to-noise ratio. We integrated the CoP-seq and Hi-C technique (Hi-CoP) for the detection of interactions of accessible chromatin regions, which represent active cis -regulatory elements in cells. We observed that the HiCoP captured the peaks in the promoters-associated enhancer regions, and the chromatin features identified by HiCoP were similar to HiChIP of histone H3K27 acetylation rather than Hi-C. HiCoP detected more promoter-enhancer (P-E), promoter-promoter (P-P), and enhancer-enhancer (E-E) interactions within 20kb-5Mb than Hi-C. Most of the loops identified by HiCoP were associated with the expressed genes. Conclusion: CoP assay can efficiently enrich open chromatin regions. When CoP assay was integrated with Hi-C assay, it provides a simple, robust, alternative technique for profiling accessible chromatin regions and chromatin conformation simultaneously.

scholarly journals Fine-mapping cellular QTLs with RASQUAL and ATAC-seq

2015 ◽  
Author(s):  
Natsuhiko Kumasaka ◽  
Andrew Knights ◽  
Daniel Gaffney
Keyword(s):  
Fine Mapping ◽  
Population Level ◽  
Regulatory Elements ◽  
Causal Variant ◽  
European Population ◽  
Open Chromatin ◽  
Gene Promoters ◽  
Genetic Associations ◽  
Functional Interpretation ◽  
Allele Specific

When cellular traits are measured using high-throughput DNA sequencing quantitative trait loci (QTLs) manifest at two levels: population level differences between individuals and allelic differences between cis-haplotypes within individuals. We present RASQUAL (Robust Allele Specific QUAntitation and quality controL), a novel statistical approach for association mapping that integrates genetic effects and robust modelling of biases in next generation sequencing (NGS) data within a single, probabilistic framework. RASQUAL substantially improves causal variant localisation and sensitivity of association detection over existing methods in RNA-seq, DNaseI-seq and ChIP-seq data. We illustrate how RASQUAL can be used to maximise association detection by generating the first map of chromatin accessibility QTLs (caQTLs) in a European population using ATAC-seq. Despite a modest sample size, we identified 2,706 independent caQTLs (FDR 10%) and illustrate how RASQUAL's improved causal variant localisation provides powerful information for fine-mapping disease-associated variants. We also map “multipeak” caQTLs, identical genetic associations found across multiple, independent open chromatin regions and illustrate how genetic signals in ATAC-seq data can be used to link distal regulatory elements with gene promoters. Our results highlight how joint modelling of population and allele-specific genetic signals can improve functional interpretation of noncoding variation.

scholarly journals A Chromatin Accessibility Atlas of the Developing Human Telencephalon

2019 ◽  
Author(s):  
Eirene Markenscoff-Papadimitriou ◽  
Sean Whalen ◽  
Pawel Przytycki ◽  
Reuben Thomas ◽  
Fadya Binyameen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
De Novo ◽  
Cell Types ◽  
Regulatory Elements ◽  
Chromatin Accessibility ◽  
Open Chromatin ◽  
Deep Layers ◽  
Health And Disease ◽  
Human Telencephalon

AbstractGene expression differs between cell types and regions within complex tissues such as the developing brain. To discover regulatory elements underlying this specificity, we generated genome-wide maps of chromatin accessibility in eleven anatomically-defined regions of the developing human telencephalon, including upper and deep layers of the prefrontal cortex. We predicted a subset of open chromatin regions (18%) that are most likely to be active enhancers, many of which are dynamic with 26% differing between early and late mid-gestation and 28% present in only one brain region. These region-specific predicted regulatory elements (pREs) are enriched proximal to genes with expression differences across regions and developmental stages and harbor distinct sequence motifs that suggest potential upstream regulators of regional and temporal transcription. We leverage this atlas to identify regulators of genes associated with autism spectrum disorder (ASD) including an enhancer of BCL11A, validated in mouse, and two functional de novo mutations in individuals with ASD in an enhancer of SLC6A1, validated in neuroblastoma cells. These applications demonstrate the utility of this atlas for decoding neurodevelopmental gene regulation in health and disease.SummaryTo discover regulatory elements driving the specificity of gene expression in different cell types and regions of the developing human brain, we generated an atlas of open chromatin from eleven dissected regions of the mid-gestation human telencephalon, including upper and deep layers of the prefrontal cortex. We identified a subset of open chromatin regions (OCRs), termed predicted regulatory elements (pREs), that are likely to function as developmental brain enhancers. pREs showed regional differences in chromatin accessibility, including many specific to one brain region, and were correlated with gene expression differences across the same regions and gestational ages. pREs allowed us to map neurodevelopmental disorder risk genes to developing telencephalic regions, and we identified three functional de novo noncoding variants in pREs that alter enhancer function. In addition, transgenic experiments in mouse validated enhancer activity for a pRE proximal to BCL11A, showing how this atlas serves as a resource for decoding neurodevelopmental gene regulation in health and disease.

scholarly journals RNA polymerase mapping in plants identifies intergenic regulatory elements enriched in causal variants

2021 ◽  
Author(s):  
Roberto Lozano ◽  
Gregory T Booth ◽  
Bilan Yonis Omar ◽  
Bo Li ◽  
Edward S Buckler ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Cell Function ◽  
Regulatory Element ◽  
Crop Improvement ◽  
Regulatory Elements ◽  
Open Chromatin ◽  
Control Of Gene Expression ◽  
Causal Variants ◽  
Divergent Transcription ◽  
Genomic Regions

Abstract Control of gene expression is fundamental at every level of cell function. Promoter-proximal pausing and divergent transcription at promoters and enhancers, which are prominent features in animals, have only been studied in a handful of research experiments in plants. PRO-Seq analysis in cassava (Manihot esculenta) identified peaks of transcriptionally engaged RNA polymerase at both the 5′ and 3′ end of genes, consistent with paused or slowly moving Polymerase. In addition, we identified divergent transcription at intergenic sites. A full genome search for bi-directional transcription using an algorithm for enhancer detection developed in mammals (dREG) identified many intergenic regulatory element (IRE) candidates. These sites showed distinct patterns of methylation and nucleotide conservation based on genomic evolutionary rate profiling (GERP). SNPs within these IRE candidates explained significantly more variation in fitness and root composition than SNPs in chromosomal segments randomly ascertained from the same intergenic distribution, strongly suggesting a functional importance of these sites. Maize GRO-Seq data showed RNA polymerase occupancy at IREs consistent with patterns in cassava. Furthermore, these IREs in maize significantly overlapped with sites previously identified on the basis of open chromatin, histone marks, and methylation, and were enriched for reported eQTL. Our results suggest that bidirectional transcription can identify intergenic genomic regions in plants that play an important role in transcription regulation and whose identification has the potential to aid crop improvement.

scholarly journals Profiling of accessible chromatin regions across multiple plant species and cell types reveals common gene regulatory principles and new control modules

2017 ◽  
Author(s):  
Kelsey A. Maher ◽  
Marko Bajic ◽  
Kaisa Kajala ◽  
Mauricio Reynoso ◽  
Germain Pauluzzi ◽  
...  
Keyword(s):  
Hair Cell ◽  
Plant Species ◽  
Stress Responses ◽  
Binding Sites ◽  
Cell Types ◽  
Regulatory Elements ◽  
Open Chromatin ◽  
Cell Type ◽  
Cell Type Specific ◽  
Accessible Chromatin

ABSTRACTThe transcriptional regulatory structure of plant genomes remains poorly defined relative to animals. It is unclear how many cis-regulatory elements exist, where these elements lie relative to promoters, and how these features are conserved across plant species. We employed the Assay for Transposase-Accessible Chromatin (ATAC-seq) in four plant species (Arabidopsis thaliana, Medicago truncatula, Solanum lycopersicum, and Oryza sativa) to delineate open chromatin regions and transcription factor (TF) binding sites across each genome. Despite 10-fold variation in intergenic space among species, the majority of open chromatin regions lie within 3 kb upstream of a transcription start site in all species. We find a common set of four TFs that appear to regulate conserved gene sets in the root tips of all four species, suggesting that TF-gene networks are generally conserved. Comparative ATAC-seq profiling of Arabidopsis root hair and non-hair cell types revealed extensive similarity as well as many cell type-specific differences. Analyzing TF binding sites in differentially accessible regions identified a MYB-driven regulatory module unique to the hair cell, which appears to control both cell fate regulators and abiotic stress responses. Our analyses revealed common regulatory principles among species and shed light on the mechanisms producing cell type-specific transcriptomes during development.

scholarly journals Disruption of c-MYC Binding and Chromosomal Looping Involving Genetic Variants Associated With Ankylosing Spondylitis Upstream of the RUNX3 Promoter

2022 ◽  
Vol 12 ◽  
Author(s):  
Carla J. Cohen ◽  
Connor Davidson ◽  
Carlo Selmi ◽  
Paul Bowness ◽  
Julian C. Knight ◽  
...  
Keyword(s):  
Ankylosing Spondylitis ◽  
Binding Site ◽  
Regulatory Elements ◽  
Published Data ◽  
Open Chromatin ◽  
Physical Interaction ◽  
Nucleotide Polymorphisms ◽  
Genetic Associations ◽  
Chromosome Conformation ◽  
Nuclear Extracts

Background: Ankylosing Spondylitis (AS) is a common form of inflammatory spinal arthritis with a complex aetiology and high heritability, involving more than 100 genetic associations. These include several AS-associated single nucleotide polymorphisms (SNPs) upstream of RUNX3, which encodes the multifunctional RUNT-related transcription factor (TF) 3. The lead associated SNP rs6600247 (p = 2.6 × 10−15) lies ∼13kb upstream of the RUNX3 promoter adjacent to a c-MYC TF binding-site. The effect of rs6600247 genotype on DNA binding and chromosome looping were investigated by electrophoretic mobility gel shift assays (EMSA), Western blotting-EMSA (WEMSA) and Chromosome Conformation Capture (3C).Results: Interrogation of ENCODE published data showed open chromatin in the region overlapping rs6600247 in primary human CD14+ monocytes, in contrast to the Jurkat T cell line or primary human T-cells. The rs6600247 AS-risk allele is predicted to specifically disrupt a c-MYC binding-site. Using a 50bp DNA probe spanning rs6600247 we consistently observed reduced binding to the AS-risk “C” allele of both purified c-MYC protein and nuclear extracts (NE) from monocyte-like U937 cells. WEMSA on U937 NE and purified c-MYC protein confirmed these differences (n = 3; p &lt; 0.05). 3C experiments demonstrated negligible interaction between the region encompassing rs6600247 and the RUNX3 promoter. A stronger interaction frequency was demonstrated between the RUNX3 promoter and the previously characterised AS-associated SNP rs4648889.Conclusion: The lead SNP rs6600247, located in an enhancer-like region upstream of the RUNX3 promoter, modulates c-MYC binding. However, the region encompassing rs6600247 has rather limited physical interaction with the promoter of RUNX3. In contrast a clear chromatin looping event between the region encompassing rs4648889 and the RUNX3 promoter was observed. These data provide further evidence for complexity in the regulatory elements upstream of the RUNX3 promoter and the involvement of RUNX3 transcriptional regulation in AS.

scholarly journals Functional Annotation of Putative Regulatory Elements at Cancer Susceptibility Loci

2014 ◽  
Vol 13s2 ◽  
pp. CIN.S13789
Author(s):  
Stephanie A. Rosse ◽  
Paul L. Auer ◽  
Christopher S. Carlson
Keyword(s):  
Cancer Susceptibility ◽  
Association Studies ◽  
Regulatory Elements ◽  
Genome Wide Association Studies ◽  
Genetic Associations ◽  
Functional Polymorphisms ◽  
Functional Variants ◽  
Genome Wide ◽  
Regulatory Variant ◽  
Causal Variants

Most cancer-associated genetic variants identified from genome-wide association studies (GWAS) do not obviously change protein structure, leading to the hypothesis that the associations are attributable to regulatory polymorphisms. Translating genetic associations into mechanistic insights can be facilitated by knowledge of the causal regulatory variant (or variants) responsible for the statistical signal. Experimental validation of candidate functional variants is onerous, making bioinformatic approaches necessary to prioritize candidates for laboratory analysis. Thus, a systematic approach for recognizing functional (and, therefore, likely causal) variants in noncoding regions is an important step toward interpreting cancer risk loci. This review provides a detailed introduction to current regulatory variant annotations, followed by an overview of how to leverage these resources to prioritize candidate functional polymorphisms in regulatory regions.

scholarly journals HiCoP, a simple and robust method for detecting interactions of regulatory regions

2020 ◽  
Author(s):  
Yan Zhang ◽  
Zhaoqiang Li ◽  
Shasha Bian ◽  
Hao Zhao ◽  
Delong Feng ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Regulatory Elements ◽  
High Signal ◽  
Open Chromatin ◽  
Regulatory Regions ◽  
Essential Information ◽  
Histone H3k27 ◽  
Development And Differentiation ◽  
Physical Interactions ◽  
Accessible Chromatin

Abstract Background: Chromatin physical interactions provide essential information for understanding the regulation of cis-elements like enhancers, promoters, and insulators in cell development and differentiation. The Hi-C assay is a technique detecting chromatin structures of the whole genome but not sensitive to interactions of regulatory elements. Several methods, like HiChIP, DNase-C, and OCEAN-C, have been developed for enriching interactions of regulatory regions, but all of them have some shortcomings. New simple, efficient, and robust methods are still in need of detecting interactions of regulatory regions. Results: We developed a new, simple, and robust assay called CoP (Column Purified chromatin) for profiling of open chromatin regions by directly purifying fragmentized crosslinked chromatin with a DNA purification column. The accessible chromatin regions, including active enhancers, promoters, and insulators, were significantly enriched in CoP chromatin. The CoP-seq assay can efficiently detect open chromatin regions, especially active promoters, with a high signal-to-noise ratio. We integrated the CoP-seq and Hi-C technique (HiCoP) for the detection of interactions of accessible chromatin regions, which represent active cis-regulatory elements in cells. We observed that the HiCoP captured the peaks in the promoters-associated enhancer regions, and the chromatin features identified by HiCoP were similar to HiChIP of histone H3K27 acetylation rather than Hi-C. HiCoP detected more promoter-enhancer (P-E), promoter-promoter (P-P), and enhancer-enhancer (E-E) interactions within 20kb-5Mb than Hi-C. Most of the loops identified by HiCoP were associated with the expressed genes. Conclusion: CoP assay can efficiently enrich open chromatin regions. When CoP assay was integrated with Hi-C assay, it provides a simple, robust, alternative technique for profiling accessible chromatin regions and chromatin conformation simultaneously.

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