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1,149 results for “transgenic”
Comprehensive characterisation of the genomic insertion site of a transgene in highly repetitive, centromeric region of Anopheles mosquitoes
<p>The availability of the genomic sequence of the malaria mosquito <em>Anopheles</em> <em>gambiae</em> has sparked in recent years the development of transgenic technologies with the potential to be used as novel tools for vector control. These technologies rely on genome editing that confers features able to affect vector capacity. This can be achieved by either reducing the mosquito population or by making mosquitoes refractory to the parasite infection. Although sophisticated molecular techniques such as those based on AttB/AttP site-specific recombination and CRISPR/Cas9 systems can lead to the integration of transgenes in specific sites of the genome, methods that allow semi-random integration are still in use due to their high efficiency; PiggyBac transposon-mediated integrations fall in this category. Characterization of the insertion site of transgenes in transgenic strains generated via PiggyBac integration can be hampered when the transgene is inserted in regions of the genome rich in repetitive sequences. Here we describe a number of techniques that were used to identify the genomic location of the transgene in a repetitive region in the <em>Anopheles gambiae</em> strain Ag(PMB)1 which was initially reported on Chromosome 3R 36D. Whilst Inverse PCR used in previous analysis was unable to distinguish between multiple genomic locations as potential insertion sites of the transgene, here we demonstrate that the use of FISH identifies clearly the integration of the transgene in a poorly annotated centromeric region of Chromosome 2R 19D. This study emphasises the need for accuracy in sequencing data for the genome of organisms of medical importance such as <em>Anopheles </em>mosquitoes. An effort to further improve reference genomes is of paramount importance to support and facilitate vector control interventions based on genome editing.</p>
Targeted sequencing of T-DNA borders in OCP1xOGC transgenic lines of Camelina
<p>Background: Genetic engineering of crop plants has been successful in transferring traits into elite lines beyond what can be achieved with breeding techniques. Introduction of transgenes originating from other species has conferred resistance to biotic and abiotic stresses, increased efficiency, and modified developmental programs. The next challenge is now to combine multiple transgenes into elite varieties via gene stacking to combine traits. Generating stable homozygous lines with multiple transgenes requires selection of segregating generations which is time consuming and labor intensive, especially if the crop is polyploid. Insertion site effects and transgene copy number are important metrics for commercialization and trait efficiency.</p> <p>Results: We have developed a simple method to identify the sites of transgene insertions using T-DNA-specific primers and high-throughput sequencing that enables identification of multiple insertion sites in the T1 generation of any crop transformed via <em>Agrobacterium</em>. We present an example using the allohexaploid oil-seed plant <em>Camelina sativa</em> to determine insertion site location of two transgenes.</p> <p>Conclusion: This new methodology enables the early selection of desirable transgene location and copy number to generate homozygous lines within two generations.</p>
Quantification of transgene expression in GSH AAVS1 with a novel CRISPR/Cas9 based approach reveals high transcriptional variation
<p>Inderbitzin, Loosli and colleagues employ a novel method to construct and extensively characterize DNA barcode libraries and apply CRISPR/Cas9 technology for targeted insertion into the safe harbor gene AAVS1 in Jurkat cells. This technique revealed high fluctuations in gene expression in AAVS1, spanning over two logs.</p>
Noscapine treatment effect in transgenic mouse model of Alzheimer's disease
<p>Cerebrovascular dysfunction and neuroinflammation play key roles in the pathophysiology of Alzheimer’s disease (AD). The kinin-kallikrein system involving bradykinin receptor has been proposed at the nexus of beta-amyloid, vascular pathology and inflammation in patients with AD and in animal models. Here, we evaluated the effect of blocking the bradykinin receptors 1 and 2 by treatment with the bradykinin antagonist noscapine on cerebrovascular dysfunction, inflammation and amyloid pathology in a transgenic mouse model of amyloidosis. Transgenic arcAβ mice, and wild-type littermates of 14 months-of-age were either treated with noscapine (3 g/L, acidified drinking water) or received drinking water as control for three months (n = 8-11 per group). Arterial spin labeling magnetic resonance imaging showed alleviated regional hypoperfusion in noscapine-treated arcAb compared to control arcAb mice. Functional magnetic resonance imaging showed mitigated reduced regional cerebral vascular reactivity in noscapine-treated arcAb compared to control arcAb mice.</p>
Scans of leaves dissected in phyllotactic order: complementation of ago7 mutant A. thaliana plants with truncated promoter transgenes
<p>This transgenic complementation experiment is described in a paper by Hoyer et al. (2019): <a href="https://doi.org/10.1002/pld3.102">https://doi.org/10.1002/pld3.102</a></p> <p>Adaxial (ad) and abaxial (ab) surfaces of rosettes were photographed in between removal of leaves, to enable checking that leaves were removed in the correct phyllotactic order.<strong> </strong>A manifest file with file SHA-1 sums is included.</p> <p>Metadata and LeafJ measurements have been made available separately, to facilitate updates -- see <a href="https://github.com/jshoyer/raspi-photo-and-leaf-scan-metadata">https://github.com/jshoyer/raspi-photo-and-leaf-scan-metadata</a>, archived as <a href="https://doi.org/10.5281/zenodo.1340636">https://doi.org/10.5281/zenodo.1340636</a></p> <p>Seed was plated and growth started on 2016-08-26. Leaves were dissected and scanned 33 and 35 days post-stratification (2016-09-28 and 30).</p>
Time-lapse photograph dataset: complementation of ago7 mutant A. thaliana plants with truncated promoter transgenes
<p>This transgenic complementation experiment is described in a paper by Hoyer et al. (2019): <a href="https://doi.org/10.1002/pld3.102">https://doi.org/10.1002/pld3.102</a></p> <p>Time-stamped photographs are provided in twelve directories by camera (twelve overlapping fields of view) and a manifest file with file SHA-1 sums is included.</p> <p>Metadata have been made available separately, to facilitate updates -- see <a href="https://github.com/jshoyer/raspi-photo-and-leaf-scan-metadata">https://github.com/jshoyer/raspi-photo-and-leaf-scan-metadata</a>, archived as <a href="https://doi.org/10.5281/zenodo.1340636">https://doi.org/10.5281/zenodo.1340636</a></p> <p>Seed was plated and growth started on 2016-08-26.</p>
Transgenic A53T mice have astrocytic -synuclein aggregates in dopamine and striatal regions
<p>Statistical analysis carried out on astrocyte quantification data derived from 6 month transgenic A53T PD mice. </p>
Transgenic A53T mice have astrocytic a-synuclein aggregates in dopamine and striatal regions
<p>Statistical analysis carried out on astrocyte quantification data derived from 6 month transgenic A53T PD mice. </p>
DNA sequences of transgenes detected via environmental DNA (raw ABI files, processed FASTA files, and reference alignments)
We demonstrate that simple, non-invasive environmental DNA (eDNA) methods can detect transgenes of genetically modified (GM) animals from terrestrial and aquatic sources in invertebrate and vertebrate systems. We detected transgenic fragments between 82-234 bp through targeted PCR amplification of environmental DNA extracted from food media of GM fruit flies (<i>Drosophila melanogaster</i>), feces, urine, and saliva of GM laboratory mice (<i>Mus musculus</i>), and aquarium water of GM tetra fish (<i>Gymnocorymbus ternetzi</i>). With rapidly growing accessibility of genome-editing technologies such as CRISPR, the prevalence and diversity of GM animals will increase dramatically. GM animals have already been released into the wild with more releases planned in the future. eDNA methods have the potential to address the critical need for sensitive, accurate, and cost-effective detection and monitoring of GM animals and their transgenes in nature.
Chronic hyperadiponectinemia induced by transgenic overexpression increases plasma exosomes without significantly improving glucose and lipid metabolism
<p><span>The fat</span><span>-derived factor, adiponectin, is considered a salutary circulating factor.</span> <span>We recently demonstrated that native adiponectin binds T-cadherin and promotes intracellular biogenesis and secretion of the exosome. Exosomes play important roles in various aspects of homeostasis, including glucose and energy metabolism. However, it remains unclear whether and how the promotion of exosome production by adiponectin <em>in vivo</em> is beneficial for glucose and lipid metabolism. In the present study, overexpression of human adiponectin in mice resulted in an increased number of circulating exosomes, but it did not significantly improve glucose metabolism, change body weights, or change triglyceride clearance under a high-fat diet. Multiple small doses of streptozotocin increased blood glucose and decreased triglyceride clearance similarly in both wild-type and transgenic mice. Thus, these results indicated that human adiponectin overexpression in mice increases plasma exosomes but does not significantly influence glucose and lipid metabolism.</span></p>
Omicron sub-lineage BA.5 infection results in attenuated pathology in hACE2 transgenic mice
<p>Omicron recently emerged sub-lineage BA.5, together with BA.4, caused a 5th wave of coronavirus disease (COVID-19) in South Africa and subsequently emerged as a predominant strain globally due to its high transmissibility. The lethality of BA.5 infection has not been studied in acute hACE2 transgenic (hACE2.Tg) mice models. Here, we investigated tissue-tropism and immuno-pathology induced by BA.5 infection in hACE2.Tg mice. Our data show that intranasal BA.5 infection in hACE2.Tg mice result in attenuated pulmonary infection and pathology with diminished COVID-19-induced clinical and pathological manifestations. BA.5, similar to Omicron (B.1.1.529), infection led to attenuated production of inflammatory cytokines, anti-viral response and effector T cell response as compared to the ancestral strain. Mice recovered from B.1.1.529 infection showed robust protection against BA.5 infection with reduced lung viral load and pathology. Our data provide insights as to why BA.5 infection escapes previous SARS-CoV-2 exposure induced-T cell immunity but may result in milder immuno-pathology and alleviated chances of re-infectivity in Omicron-recovered individuals.</p>
Computationally defined and in vitro validated putative genomic safe harbour loci for transgene expression in human cells
<p>Selection of the target site is an inherent question for any project aiming for directed transgene integration. Genomic safe harbour (GSH) loci have been proposed as safe sites in the human genome for transgene integration. Although several sites have been characterised for transgene integration in the literature, most of these do not meet criteria set out for a GSH, and the limited set that do have not been characterised extensively. Here, we conducted a computational analysis using publicly available data to identify 25 unique putative GSH loci that reside in active chromosomal compartments. We validated stable transgene expression and minimal disruption of the native transcriptome in three GSH sites <em>in vitro</em> using human embryonic stem cells (hESCs) and their differentiated progeny. Furthermore, for easily targeted transgene expression, we have engineered constitutive landing pad expression constructs into the three validated GSH in hESCs.</p>
Omicron sub-lineage BA.5 infection results in attenuated pathology in hACE2 transgenic mice
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Data for: Phytochemical shift from condensed tannins to flavonoids in transgenic Betula pendula decreases consumption and growth but improves growth efficiency of Epirrita autumnata larvae
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DNA sequences of transgenes detected via environmental DNA (raw ABI files, processed FASTA files, and reference alignments)
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Data from: Establishment of transgenic <em>Drosophila suzukii</em> lines that express <em>phiC31</em> integrase and carry the <em>sepia</em> gene as a marker for transformation
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Chronic hyperadiponectinemia induced by transgenic overexpression increases plasma exosomes without significantly improving glucose and lipid metabolism
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Early ultrasonic vocalization deficits and related thyroarytenoid muscle pathology in the transgenic TgF344-AD rat model of Alzheimer’s disease
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Data from: Evaluating transgenic Darling 54 American chestnuts for reintroduction: Insights from survivorship, growth, and respiration in a common garden
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Data from: Detecting the population dynamics of an autosomal sex-ratio distorter transgene in malaria vector mosquitoes
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Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.