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Andre, C.

Publications and source records attributed to Andre, C..

At least 19 recordsLinked to original sources

Alterations to the yeast chromatin landscape during hypoxia

When cells encounter low oxygen conditions, signaling events target key transcription factors to reprogram gene expression to promote cell survival and adaptation under this challenging environmental stress. The substantial metabolic changes that occur in hypoxia also drive changes to the chromatin environment due to altered cofactor availability and direct oxygen sensitivity of chromatin-modifying enzymes, among other factors. Given the links between hypoxic cellular environments and numerous pathophysiological processes, including tumor progression and metastasis, cardiovascular disorders, and aberrant development, it is critical to understand the chromatin states and chromatin-based regulatory mechanisms associated with cellular responses to hypoxia. Here, we used the model eukaryote Saccharomyces cerevisiae to interrogate the requirement for key transcription factors and chromatin regulators in survival during hypoxia and we assessed the distribution of major histone modifications associated with the response to stress, H3K9 acetylation and H3K4 methylation, throughout the genome during hypoxia. Our results show that only a small number of chromatin modifiers in yeast contribute to survival in hypoxia; however, there are substantial changes to the abundance and distribution of both H3K9ac and H3K4me3 during hypoxic growth. Specifically, H3K9ac levels are reduced throughout the genome though still maintain association with transcriptionally active genes. H3K4me3 has increased abundance genome-wide and shows a wider distribution with greater abundance in coding sequences particularly of genes upregulated in hypoxic conditions. Altogether, our data demonstrate substantial changes to the chromatin landscape in hypoxic conditions in the budding yeast model, providing key insights into physiological chromatin changes caused by limiting oxygen in the environment.

molecular biology↗

Establishing a baseline for standardised genetic monitoring of Atlantic cod (Gadus morhua) in Sweden

Protecting populations and genetic diversity within them is critical to conserving the resilience and adaptive potential of species. Fisheries management has long had the ambition of managing species at the population level, but mainly define "fish stocks" based on geographical limits, which can lead to overfishing of sensitive populations in areas where many different populations coexist. Modern genetic methods are now sufficiently cost-effective, fast, and accurate to be integrated into fisheries management, enabling genetic identification and monitoring of fish populations. Here, we establish a genetic baseline for the commercially important fish Atlantic cod (Gadus morhua) in the waters surrounding Sweden, by using standardised sampling procedures and developing a genetic panel of 4000 single nucleotide polymorphisms (SNPs) for cost-effective assignment of population-of-origin and inversion genotypes. Using the SNP panel, we resolve the geographical distribution of three genetically distinct cod populations in the region: offshore, coastal/Western Baltic, and Eastern Baltic cod. While there is considerable spatial overlap between the three populations, they are genetically differentiated across the entire genome, as well as in genomic regions associated with chromosomal inversions. In addition, heterozygosity and effective population size estimates suggest differences in genetic diversity and rates of genetic erosion, underscoring the need to monitor the genetic diversity within each population separately. Repeating this methodology across years provides a first suggestion for establishing spatiotemporally resolved genetic monitoring of Atlantic cod in Sweden - simultaneously accounting for both population structure within the species and the genetic diversity within populations.

genomics↗

Alpha hemolysin polymorphisms in methicillin-resistant Staphylococcus aureus clinical isolates regulate ADAM10-dependent neutrophil IL-1 beta secretion

Staphylococcus aureus -hemolysin (Hla) is a major virulence factor that utilizes cell surface ADAM10 to oligomerize and form a functional heptameric pore. We show here that Hla from strain USA300 is required to induce IL-1{beta} secretion by neutrophils and to cause severe corneal disease in mice. We also demonstrate that in contrast to USA300 and other clonal complex 8 (CC8) methicillin resistant S. aureus (MRSA) isolated from the skin, CC5 Hla from corneas of infected patients have single nucleotide polymorphisms (SNP) that result in two amino acid substitutions, D208E (Asp-Glu) and I275T (Ile-Thr). Structural modeling predicts CC5 Hla self-assembly and altered binding to ADAM10 that is distinct from CC8 Hla. The ADAM10 inhibitor GI254023X blocked neutrophil IL-1{beta} secretion induced by Hla-expressing CC8, but not by CC5 conditioned media, indicating that these Hla polymorphisms play an important role in Hla receptor binding and neutrophil IL-1{beta} secretion, and affect corneal disease severity.

microbiology↗

Facilitating genome annotation using ANNEXA and long-read RNA sequencing

With the advent of complete genome assemblies, genome annotation has become essential for the functional interpretation of genomic data. Long-read RNA sequencing (LR-RNAseq) technologies have significantly improved transcriptome annotation by enabling full-length transcript reconstruction for both coding and non-coding RNAs. However, challenges such as transcript fragmentation and incomplete isoform representation persist, highlighting the need for robust quality control (QC) strategies. This study presents ANNEXA, a pipeline designed to enhance genome annotation using LR-RNAseq data while also providing QC for reconstructed genes and transcripts. ANNEXA integrates two transcriptome reconstruction tools, StringTie2 and Bambu, applying stringent filtering criteria to improve annotation accuracy. It also incorporates deep learning models to evaluate transcription start sites (TSSs) and employs the tool FEELnc for the systematic annotation of long non-coding RNAs (lncRNAs). Additionally, the pipeline offers intuitive visualisations for comparative analyses of coding and non-coding repertoires. Benchmarking against multiple reference annotations revealed distinct patterns of sensitivity and precision for both known and novel genes and transcripts and mRNAs and lncRNAs. To demonstrate its utility, ANNEXA was applied in a comparative oncology study involving LR-RNAseq of two human and eight canine cancer cell lines. The pipeline successfully identified novel genes and transcripts across species, expanding the catalog of protein-coding and lncRNA annotations in both species. Implemented in Nextflow for scalability and reproducibility, ANNEXA is available as an open-source tool: https://github.com/IGDRion/ANNEXA.

bioinformatics↗

Wafer-scale integration of alpha-quartz thin films towards super high frequency piezoelectric bioNEMS for arbovirus detection

Micro and nanoelectromechanical systems (MEMS/NEMS), especially piezoelectric resonators, offer a promising strategy for the manufacturing of point-of-care devices providing rapid, sensitive, and field-deployable tests with minimal user training for the diagnostic of viral infections. High-frequency (HF) MEMS/NEMS have the potential for ultrasensible mass-loading devices. Yet, their use for biomedical applications requires challenging manufacturing qualities. Here, we develop a large-scale chemical integration of epitaxial -quartz (100) thin films on silicon wafers up to 4-inches. This methodology allows the microfabrication of wafer-scale piezoelectric -quartz/silicon bioMEMS using a recognition layer capable of selectively detecting emerging arboviruses over other viral loads. Using contact-free vibrometry, we show a mass sensitivity of the bioMEMS device of 22.4 pg/Hz in liquid conditions and a Chikungunya virus limit of detection of 9 ng/ml. To reach piezoelectric transduction for compact quartz sensor devices, we develop NEMS resonators at super HF, i.e., 17.8 GHz with a quality factor of 280 which represents a QxF product of 4.98{middle dot}1012. These -quartz NEMS can reach thicknesses between 100 and 800 nm and lateral dimensions up to 9 mm2. Our work opens the door for cost-efficient single-chip epitaxial piezoelectric -quartz/Si ultrasensitive NEMS sensors manufactured exclusively by soft-chemistry for biomedical applications and many other fields.

bioengineering↗

Resting-state functional connectivity and fast spindle temporal organization contribute to episodic memory consolidation in healthy aging

Episodic memory consolidation relies on the functional specialization of brain networks and sleep quality, both of which are affected by aging. Functional connectivity during wakefulness is crucial to support the integration of newly acquired information into memory networks. Additionally, the temporal dynamics of sleep spindles facilitates overnight memory consolidation by promoting hippocampal replay and integration of memories within neocortical structures. This study aimed at exploring how resting-state functional connectivity during wakefulness contributes to sleep-dependent memory consolidation in aging, and whether spindles clustered in trains modulates this relationship. Forty-two healthy older adults (68.82 {+/-} 3.03 years), enrolled in the Age-Well clinical trial, were included. Sleep-dependent memory consolidation was assessed using a visuo-spatial memory task performed before and after a polysomnography night. Resting-state functional connectivity data were analyzed using graph theory applied to the whole brain, specific brain networks and the hippocampus. Lower limbic network integration and higher centrality of the anterior hippocampus were associated with better memory consolidation. Spindle trains modulated these effects, such that older participants with longer spindle trains exhibited a stronger negative association between limbic network integration and memory consolidation. These results indicate that lower functional specialization at rest is associated with weaker memory consolidation during sleep. This aligns with the dedifferentiation hypothesis, which posits that aging is associated with reduced brain specificity, leading to less efficient cognitive functioning. These findings reveal a novel mechanism linking daytime brain network organization and sleep-dependent memory consolidation, and suggest that targeting spindle dynamics could help preserve cognitive functioning in aging.

neuroscience↗

Connectivity and population structure in a marginal sea - a review

The current biodiversity crisis calls for conservation measures that limit the negative human impact on important habitats and sensitive wild populations. To effectively protect biodiversity at all levels, including intraspecific diversity, conservation measures should be aligned with the connectivity and genetic structure of wild populations. In this review, we synthesise scientific literature on the connectivity and population structure of marine species in the Skagerrak - a marginal sea in the northeast Atlantic Ocean. We discuss the results in relation to the current management practices in the region, as well as the general transferability of our findings. The Skagerrak is one of the most intensively studied regions within this research field, and our findings show that the overall connectivity with adjacent seas is high, but asymmetric, for most species. Simultaneously, most species have populations in the Skagerrak distinct from both each other, and those in adjacent seas. Most of this population structure is associated with the convoluted Skagerrak coastline - population structure is common both among coastal populations and between coastal and offshore populations. In many mobile species, multiple populations are temporally sympatric in certain areas, but retain their genetic divergence through natal homing or other barriers to gene flow. The presence of population structure despite high connectivity is a challenge for area-based protection measures, and calls for temporally flexible management that also monitors intraspecific genetic diversity on multiple timescales.

ecology↗

Analysis of in-patient evolution of Escherichia coli reveals potential links to relapse of bone and joint infections

Bone and joint infections (BJIs) are difficult to treat and affect a growing number of patients, in which relapses are observed in 10-20% of the case. These relapses, which call for prolonged antibiotic treatment and increase the risk of emergence of resistance, may originate from ill understood adaptation of the pathogen to the host. Here, we studied three pairs of Escherichia coli strains corresponding to three cases of BJIs and their relapse to better understand in-patient adaptation. Whole genome comparison presented evidence for positive selection with prevalence of non-synonymous and loss of function mutations. Phenotypic characterization showed that biofilm formation capacity was not modified, contrary to what is usually described in such relapse cases. Although virulence was not modified, we identified the loss of two virulence factors (namely an AFA afimbrial adhesin and a YadA-like adhesin) contributing to immune system evasion in one of the studied relapse strain. Other selected strategies likely helped the relapse strains to outcompete competitors through global growth optimization and colicin production. This work highlights the variety of strategies allowing in-patient adaptation in BJIs.

microbiology↗

A single base pair duplication in SLC33A1 gene causes fetal losses and neonatal lethality in Manech Tete Rousse dairy sheep

Recently, we evidenced that the Manech Tete Rousse (MTR) deficient homozygous haplotype 2 (MTRDHH2) was likely to harbor a recessive lethal variant in ovine. In the present study, we fine mapped this region by analyzing the whole genome sequence of five MTRDHH2 heterozygous carriers compared to 95 sequences of non-carrier animals from MTR and others ovine breeds. We successfully identified a single base pair duplication in the SLC33A1 gene, resulting in a frameshift leading to a premature stop codon (p.Arg246Alafs*3). SLC33A1 acts as a transmembrane transporter of acetyl-Coenzyme A, essential for cellular metabolism. In order to assess for the lethal phenotype in homozygous MTR sheep, we generated at-risk matings by artificial insemination (AI) between rams and ewes heterozygous for the SLC33A1 variant named SLC33A1_dupG. Gestation status was checked 15 days post-AI by a molecular test from blood expression of the interferon Tau-stimulated MX1 gene, and by ultrasonography performed between 45 days and 60 days post-AI. Based on ultrasonography, the AI success was reduced by 12% compared to safe matings suggesting embryonic/fetal losses further confirmed by the molecular test based on MX1 differential expression. Forty-nine lambs were born from at-risk matings with a mortality rate of 34.7% observed before weaning. Homozygous SLC33A1_dupG lambs contributed to 47% of this mortality occurring mainly in the first five days after lambing with no obvious clinical signs. Thus, an appropriate management of SLC33A1_dupG (allele frequency of 0.04) in the MTR selection scheme should increase the overall fertility and lamb survival.

genomics↗

Homozygous haplotype deficiency in Manech Tete Rousse dairy sheep revealed a nonsense variant in MMUT gene affecting newborn lamb viability

Recessive deleterious variants are known to segregate in livestock populations as in human, and some may cause lethality when homozygous. By scanning the genome of 6,845 Manech Tete Rousse dairy sheep using phased 50k SNP genotypes and pedigree data, we searched for deficiency in homozygous haplotype (DHH). Five Manech Tete Rousse deficient homozygous haplotypes (MTRDHH1 to 5) were identified with a homozygous deficiency ranging from 84% to 100%. These haplotypes are located on OAR1 (MTRDHH2 and 3), OAR10 (MTRDHH4), OAR13 (MTRDHH5) and OAR20 (MTRDHH1), and have frequencies ranging from 7.8% to 16.6%. When comparing at-risk mating between DHH carriers to safe mating between non-carriers, two DHH (MTRDHH1 and 2) showed significant effects on decreasing artificial insemination success and/or increasing stillbirth rate. We particularly investigated the MTRDHH1 haplotype highly increasing stillbirth rate, and we identified a single nucleotide variant (SNV) inducing a premature stop codon (p.Gln409*) in the MMUT gene (methylmalonyl-CoA mutase) by using a whole genome sequencing (WGS) approach. We generated homozygous lambs for the MMUT mutation by oriented mating, and most of them died within the first 24h after birth without any obvious clinical defect. RT-qPCR and western blotting performed on post-mortem liver and kidney biological samples showed a decreased expression of MMUT mRNA in the liver and absence of a full-length MMUT protein in mutated homozygous lambs. In parallel, MTRDHH4 and MTRDHH5 showed partial association with variants in RXFP2 and ASIP genes, respectively, already known to control horned/polled and coat color phenotypes in sheep, two morphological traits accounting in the MTR breed standard. Further investigations are needed to identified the supposed recessive deleterious variant hosted by MTRDHH2 and MTRDHH3. Anyway, an appropriate management of these haplotypes/variants in the MTR dairy sheep selection program should increase the overall fertility and lamb survival. Author SummaryIn this article, we used reverse genetics screen in ovine using large genotype data available in the framework of genomic selection program in Manech Tete Rousse dairy sheep. We identified five genomic regions with a highly significant deficit in homozygous animal. These regions are thus supposed to host recessive deleterious mutations. In one of these genomic regions, we identified a nonsense mutation in MMUT that alters the functioning of this essential gene of cell metabolism, causing perinatal mortality of homozygous lambs. In this work, we also identified other regions possibly associated with morphological appearance part of the breed standard such as polledness and coat color. Increasing knowledge in these genomic regions will help the future genetic management of the Manech Tete Rousse breed, particularly to reduce lamb mortality.

genomics↗

Mixed origin of juvenile Atlantic cod (Gadus morhua) along the Swedish west coast

Cryptic population structure in exploited fish species poses a major challenge for fisheries management. Atlantic cod (Gadus morhua) is a species in which the presence of sympatric ecotypes has been known for a long time, for instance off the coast of Northern Norway. More recently, two sympatric ecotypes of cod have also been documented in the Skagerrak and Kattegat; one ecotype is of an apparent offshore origin and undertakes spawning migrations to the North Sea, and the other is resident at the coast throughout its life. However, their relative contributions of juveniles to the Swedish west coast remain poorly understood. The lack of adult cod along the Skagerrak and Kattegat coasts in recent years has led to the hypothesis that the offshore ecotype is the main source of juveniles to the area, but recent studies have shown large proportions of coastal cod inside Norwegian Skagerrak fjords. In this study, juvenile cod were collected at a high spatial resolution along the Swedish west coast, and genetically assigned to each of the two ecotypes. The results reveal that there is a considerable proportion of juvenile coastal cod in the southern Kattegat, Oresund, and in inshore Swedish Skagerrak, but that the offshore ecotype dominates in offshore areas. Model selection suggests that differences in bottom depth, rather than distance from the open sea, may explain the heterogenous spatial distribution of the two ecotypes. In addition, the two ecotypes displayed differences at loci known to be associated with environmental adaptation, suggesting that their spatial distribution is maintained by natural selection in response to specific environmental conditions.

genomics↗

Serial depletion of Baltic herring since the Viking Age revealed by ancient DNA and population genomics

Marine resource consumption has been a key component in European diet and culture since the Middle Ages, when fish consumption increased dramatically. Yet, the early origins of marine industries and the long-term ecological consequences of historical and contemporary fisheries remain debated. The Baltic Sea was home to the first "industrial" fishery [~]800 years ago targeting the Baltic herring, a species that is still economically and culturally important today. We combine modern whole genome data with ancient DNA (aDNA) to identify the first known long-distance herring trade in the region, illustrating that large-scale fish trade began during the Viking Age. We resolve population structure within the Baltic and observe demographic independence for four local herring stocks over at least 200 generations. It has been suggested that overfishing at Oresund in the 16th century resulted in a demographic shift from autumn-spawning to spring-spawning herring dominance in the Baltic. We show that while the Oresund fishery had a negative impact on the western Baltic herring stock, the existence of autumn-spawning refugia in other regions of the Baltic delayed the demographic shift to spring spawning dominance until the 20th century. Importantly, modelling demographic trajectories over time, we identify a consistent pattern of serial depletion within the Baltic that is associated with changes in fishing pressure and climate, and conclude that herring exploitation at both historical and recent intensities is not sustainable. Our results highlight the complex and enduring impacts humans have had on the marine environment well before the industrial era.

ecology↗

Global, regional, and cryptic population structure in a high gene-flow transatlantic fish

1Lumpfish (Cyclopterus lumpus) is a transatlantic marine fish displaying large population sizes and a high potential for dispersal and gene-flow. These features are expected to result in weak population structure. Here, we investigated population genetic structure of lumpfish throughout its natural distribution in the North Atlantic using two approaches: I) 4393 genome wide SNPs and 95 individuals from 10 locations, and II) 139 discriminatory SNPs and 1,669 individuals from 40 locations. Both approaches identified extensive population genetic structuring with a major split between the East and West Atlantic and a distinct Baltic Sea population, as well as further differentiation of lumpfish from the English Channel, Iceland, and Greenland. The targeted discriminatory loci displayed [~]2-5 times higher divergence than the genome wide approach, revealing further evidence of local population substructures. Lumpfish from Isfjorden in Svalbard were highly distinct but resembled most fish from Greenland. The Kattegat area in the Baltic transition zone, formed a previously undescribed distinct genetic group. Also, further subdivision was detected within North America, Iceland, West Greenland, Barents Sea, and Norway. Although lumpfish have considerable potential for dispersal and gene-flow, the observed high levels of population structuring throughout the Atlantic suggests that this species displays natal homing and potentially local populations with adaptive differences. This fine-scale population structure calls for consideration when defining management units for lumpfish fishing stocks and in decisions related to sourcing and moving lumpfish for farming and cleaner fish use.

evolutionary biology↗

Finding coarse and fine scale population structure in a coastal species: population demographics meets genomics

Population genetic studies often focus on patterns at a regional scale and use spatially aggregated samples to draw inferences about population structure and drivers, potentially masking ecologically relevant population sub-structure and dynamics. In this study we use a multidisciplinary approach combining genomic, demographic, and habitat data with an oceanographic particle drift model, to unravel the patterns of genetic structure at different scales in the black goby (Gobius niger) along the Norwegian coast. Using a high-density sampling protocol, we observed restricted gene flow both at a surprisingly fine (kms) and large (100s km) scale. Our results showed a pattern of isolation by distance related to the level of exposure along the Skagerrak coast, where sheltered sampling stations had an overall level of genetic divergence about three times higher (FST =0.0046) than levels observed among exposed samples (FST =0.0015). These results were corroborated by demographic analyses which showed that population-fluctuations decrease in synchrony with distance at much smaller scales for sheltered samples (20 km) than for exposed sites (80 km), suggesting higher population connectivity among exposed sites. We also found a pronounced genetic discontinuity between populations along the Norwegian west and east coasts, with a sharp "break" around the southern tip of Norway, likely driven both by lack of habitat and by oceanographic features.

genomics↗

A metagenomic analysis of the wrackbed microbiome indicates a phylogeographic break along the North Sea - Baltic Sea transition zone

Sandy beaches are biogeochemical hotspots that bridge marine and terrestrial ecosystems via the transfer of marine organic matter, such as seaweed (termed wrack). A keystone of this unique ecosystem is the microbial community, which helps to degrade wrack and re-mineralize nutrients. However, little is known about the wrackbed microbiome, its composition, trophic ecology, or how it varies over time and space. Here we characterize the wrackbed microbiome as well as the microbiome of a primary consumer, the seaweed fly Coelopa frigida, and examine how they change along one of the most studied ecological gradients in the world, the transition from the marine North Sea to the brackish Baltic Sea. We found that polysaccharide degraders dominated both the wrackbed and seaweed fly microbiomes but there were still consistent differences between wrackbed and fly samples. Furthermore, we observed a shift in both microbial communities and functionality between the North and Baltic Sea. These shifts were mostly due to changes in the frequency of different groups of known polysaccharide degraders (Proteobacteria and Bacteroidota). We hypothesize that microbes were selected for their abilities to degrade different polysaccharides corresponding to a shift in polysaccharide content in the seaweed communities of the North vs. Baltic Sea. Our results reveal the complexities of both the wracked microbial community, with different groups specialized to different roles, and the cascading trophic consequences of shifts in the near shore algal community.

genomics↗

Canine oral melanoma genomic and transcriptomic study defines two molecular subgroups with different therapeutical targets

Mucosal melanoma (MM) is a rare and aggressive clinical cancer that occurs mostly in the head, neck, and anogenital regions. Despite recent advances in genetics and the development of revolutionary treatments, such as immunotherapy, the prognosis for MM remains poor. Canine MM shares several clinical, histological, and genetic features with its human counterpart, offering a relevant spontaneous and immunocompetent model to decipher the genetic bases and explore treatment options for human MM. We performed an integrative genomic and transcriptomic analysis of 32 canine MM samples, which allowed us to identify two molecular subgroups differing in microenvironment and structural variant (SV) content. The overexpression of genes related to the microenvironment and T-cell response was associated with tumors harboring a lower content of structural variants, whereas the overexpression of pigmentation-related pathways and oncogenes such as TERT were associated with a high SV burden. To detail the SVs, especially those with focal amplifications, whole-genome sequencing was performed on four canine MM cell lines. We showed that focal amplifications characterized complex chromosomal rearrangements targeting oncogenes such as MDM2 or CDK4 and a recurrently amplified region on canine chromosome 30, comprising the genes TRPM7, GABPB1, USP8, and SPPL2A, were candidate oncogenes for MM. We showed that the copy numbers of these genes were significantly correlated with their expression levels. Finally, we demonstrated that the genes TRPM7, GABPB1, and SPPL2A play a role in cell proliferation; thus, these may be considered new candidate oncogenes for human MM. Our findings suggest the existence of two MM molecular subgroups that may benefit from dedicated therapies, such as immune checkpoint inhibitors or targeted therapies. These results illustrate the relevance of dog models for deciphering genetic mechanisms in spontaneous MM, along with the potential to screen for efficient targeted therapies for rare and aggressive cancers in humans.

cancer biology↗

The tree that hides the forest: identification of common predisposing loci in several hematopoietic cancers and several dog breeds.

Histiocytic sarcoma (HS) is a rare but aggressive cancer in humans and dogs. The spontaneous canine model, with the clinical, epidemiological and histological similarities with human HS and specific breed predispositions, is a unique model/opportunity to unravel the genetic bases of this cancer. In this study, we aimed to identify germline risk factors associated with the development of HS in canine predisposed breeds. We used a methodology that combined several genome-wide association studies in a multi-breed and multi-cancer approach, as well as targeted next generation sequencing, and imputation combining several breeds (Bernese mountain dog, Rottweiler, flat coated retriever and golden retriever) and three haematopoietic cancers (HS, lymphoma and mast cell tumor). Results showed that we not only refined the previously identified HS risk CDKN2A locus but we identified new loci on canine chromosomes 2, 5, 12, 14, 20, 26 and X. Capture and targeted sequencing of specific loci pointed towards the existence of regulatory variants in non coding regions and/or methylation mechanisms linked to risk haplotypes, leading to strong cancer predispositions in specific dog breeds. Our results showed that these canine cancer predisposing loci appear to be due to the additive effect of several risk haplotype involved also in other haematopoietic cancers such lymphoma or mast cell tumor, illustrating the pleiotropic nature of these canine cancer loci as observed in human oncology, thus reinforcing the interest of predisposed dog breeds to study cancer initiation and progression.

genetics↗

Detecting genetic variation and base modifications together in the same single molecules of DNA and RNA at base pair resolution using a magnetic tweezer platform.

Accurate decoding of nucleic acid variation is important to understand the complexity and regulation of genome function. Here we introduce a single-molecule platform based on magnetic tweezer (MT) technology that can identify and map the positions of sequence variation and multiple base modifications together in the same single molecules of DNA or RNA at single base resolution. Using synthetic templates, we demonstrate that our method can distinguish the most common epigenetic marks on DNA and RNA with high sensitivity, specificity and precision. We also developed a highly specific CRISPR-Cas enrichment strategy to target genomic regions in native DNA without amplification. We then used this method to enrich native DNA from E. coli and characterized the differential levels of adenine and cytosine base modifications together in molecules of up to 5 kb in length. Finally, we enriched the 5 UTR of FMR1 from cells derived from a Fragile X carrier and precisely measured the repeat expansion length and methylation status of each molecule. These results demonstrate that our platform can detect a variety of genetic, epigenetic and base modification changes concomitantly within the same single molecules.

genomics↗