transgene techniques

Shiva Salsabilian, Elena Bibineyshvili, David J Margolis, Laleh Najafizadeh
Widefield optical imaging enables longitudinal capturing of the large-scale network activity of the cerebral cortex in awake behaving animals. Utilizing this technique, we investigate how the brain's functional network structure is altered following injury. Cortical activity in transgenic calcium reporter mice expressing GCaMP6s is recorded in two sessions, prior and after inducing the injury. As a quantitative network measure, "communicability" which is indicative of how easily the information flows between nodes in a network via both direct and indirect paths, is considered...
July 2019: Conference Proceedings: Annual International Conference of the IEEE Engineering in Medicine and Biology Society
Nicolae Herndon, Jennifer Shelton, Lizzy Gerischer, Panos Ioannidis, Maria Ninova, Jürgen Dönitz, Robert M Waterhouse, Chun Liang, Carsten Damm, Janna Siemanowski, Peter Kitzmann, Julia Ulrich, Stefan Dippel, Georg Oberhofer, Yonggang Hu, Jonas Schwirz, Magdalena Schacht, Sabrina Lehmann, Alice Montino, Nico Posnien, Daniela Gurska, Thorsten Horn, Jan Seibert, Iris M Vargas Jentzsch, Kristen A Panfilio, Jianwei Li, Ernst A Wimmer, Dominik Stappert, Siegfried Roth, Reinhard Schröder, Yoonseong Park, Michael Schoppmeier, Ho-Ryun Chung, Martin Klingler, Sebastian Kittelmann, Markus Friedrich, Rui Chen, Boran Altincicek, Andreas Vilcinskas, Evgeny Zdobnov, Sam Griffiths-Jones, Matthew Ronshaugen, Mario Stanke, Sue J Brown, Gregor Bucher
BACKGROUND: The red flour beetle Tribolium castaneum has emerged as an important model organism for the study of gene function in development and physiology, for ecological and evolutionary genomics, for pest control and a plethora of other topics. RNA interference (RNAi), transgenesis and genome editing are well established and the resources for genome-wide RNAi screening have become available in this model. All these techniques depend on a high quality genome assembly and precise gene models...
January 14, 2020: BMC Genomics
Pablo Nakagawa, Javier Gomez, Justin L Grobe, Curt D Sigmund
PURPOSE OF THE REVIEW: The main goal of this article is to discuss how the development of state-of-the-art technology has made it possible to address fundamental questions related to how the renin-angiotensin system (RAS) operates within the brain from the neurophysiological and molecular perspective. RECENT FINDINGS: The existence of the brain RAS remains surprisingly controversial. New sensitive in situ hybridization techniques and novel transgenic animals expressing reporter genes have provided pivotal information of the expression of RAS genes within the brain...
January 10, 2020: Current Hypertension Reports
Paul D Fraser, Asaph Aharoni, Robert D Hall, Sanwen Huang, James J Giovannoni, Uwe Sonnewald, Alisdair R Fernie
Gene editing techniques are currently revolutionizing biology allowing far greater precision than previous mutagenic and transgenic approaches. They are becoming applicable to a wide range of plant species and biological processes. Gene editing can rapidly improve a range of crop traits including disease resistance, abiotic stress tolerance, yield, nutritional quality and additional consumer traits. Unlike transgenic approaches, however, it is not facile to forensically detect gene-editing events at the molecular level, as no foreign DNA exists in the elite line...
January 10, 2020: Plant Journal
Hidetaka Kioka, Hisakazu Kato, Takeshi Fujita, Yoshihiro Asano, Yasunori Shintani, Satoru Yamazaki, Osamu Tsukamoto, Hiromi Imamura, Mikihiko Kogo, Masafumi Kitakaze, Yasushi Sakata, Seiji Takashima
Most eukaryotic cells generate adenosine triphosphate (ATP) through the oxidative phosphorylation system (OXPHOS) to support cellular activities. In cultured cell-based experiments, we recently identified the hypoxia-inducible protein G0/G1 switch gene 2 (G0s2) as a positive regulator of OXPHOS, and showed that G0s2 protects cultured cardiomyocytes from hypoxia. In this study, we examined the in vivo protective role of G0s2 against hypoxia by generating both loss-of-function and gain-of-function models of g0s2 in zebrafish...
January 8, 2020: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Alexander D Wykes, Sherie Ma, Ross A D Bathgate, Andrew L Gundlach
Modern neuroscience utilizes transgenic techniques extensively to study the activity and function of brain neural networks. A key feature of this approach is its compatibility with molecular methods for selective transgene expression in neuronal circuits of interest. Until now, such targeted transgenic approaches have not been applied to the extensive circuitry involving the neuropeptide, relaxin-3. Pharmacological and gene knock-out studies have revealed relaxin-3 signalling modulates interrelated behaviours and cognitive processes, including stress and anxiety, food and alcohol consumption, and spatial and social memory, highlighting the potential of this system as a therapeutic target...
June 2020: IBRO Reports
Ghulam Raza, Mohan B Singh, Prem L Bhalla
The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration...
December 25, 2019: Plants (Basel, Switzerland)
Xuechao Fei, Yan Yu, Yalan Di, Li Ai, Dandan Yao, Shangying Bai, Shengjie Zhao, Jihui Lyu, Xiang Cai, Rongqiao He, Zhiqian Tong
BACKGROUND: Coenzyme Q10 (CoQ10) supplementation can improve cognition in patients with Alzheimer's disease (AD) and AD transgenic model mice. To ameliorate the discomfort that patients with AD suffer after several blood extractions, a non-invasive method for detecting urine CoQ10 levels needs to be established. METHODS: Here, we developed a new technique of fluorescence spectrophotometry with ethyl cyanoacetate (FS-ECA), on the basis of the principle that the chemical derivative obtained from the interaction between CoQ10 and ECA was detected by a fluorescence detector at λex/em  = 450/515 nm...
December 25, 2019: Journal of Clinical Laboratory Analysis
Siyu Liu, Tatsuya Fukumoto, Patrizia Gena, Peng Feng, Qi Sun, Qiang Li, Tadashi Matsumoto, Toshiyuki Kaneko, Hang Zhang, Yao Zhang, Shihua Zhong, Weizhong Zeng, Maki Katsuhara, Yoshichika Kitagawa, Aoxue Wang, Giuseppe Calamita, Xiaodong Ding
Plasma membrane intrinsic proteins (PIPs) are known to be major facilitators of movement of a number of substrates across cell membranes. Here, from a drought-resistant rice cultivar, we isolated an OsPIP1;3 gene SNP which is mostly expressed in rice roots, and is strongly responsive to drought stress. Immunocytochemistry showed that OsPIP1;3 majorly accumulated on the proximal end of the endodermis and the cell surface around xylem. Expression of GFP-OsPIP1;3 alone in Xenopus oocytes or rice protoplasts showed OsPIP1;3 mislocalization in ER-like neighborhood, whereas co-expression of OsPIP2;2 recruited OsPIP1;3 to the plasma membrane and led to a significant enhancement of water permeability in oocytes...
December 23, 2019: Plant Journal
Katherine J Zhu, Lauren M Aiani, Nigel P Pedersen
Mouse models are beneficial to understanding neural networks given a wide array of transgenic mice and cell-selective techniques. However, instrumentation of mice for neurophysiological studies is difficult. Often surgery is prolonged with experimental error arising from non-concurrent and variable implantations. While similar approaches with microelectrodes have been used in rats before, and related approaches exist for targeting tetrodes to one brain region, we do not know of similar head-plates for mice, nor a strictly source-code and easily reconfigurable approach...
December 20, 2019: Journal of Neuroscience Methods
Sharon C Yates, Nicolaas E Groeneboom, Christopher Coello, Stefan F Lichtenthaler, Peer-Hendrik Kuhn, Hans-Ulrich Demuth, Maike Hartlage-Rübsamen, Steffen Roßner, Trygve Leergaard, Anna Kreshuk, Maja A Puchades, Jan G Bjaalie
Transgenic animal models are invaluable research tools for elucidating the pathways and mechanisms involved in the development of neurodegenerative diseases. Mechanistic clues can be revealed by applying labelling techniques such as immunohistochemistry or in situ hybridisation to brain tissue sections. Precision in both assigning anatomical location to the sections and quantifying labelled features is crucial for output validity, with a stereological approach or image-based feature extraction typically used...
2019: Frontiers in Neuroinformatics
Kyle B Fischer, Hannah K Collins, Edward M Callaway
In combination with transgenic mouse lines expressing Cre or Flp recombinases in defined cell types, recombinase-dependent adeno-associated viruses (AAVs) have become the tool of choice for localized cell-type-targeted gene expression. Unfortunately, applications of this technique when expressing highly sensitive transgenes are impeded by off-target, or "leak" expression, from recombinase-dependent AAVs. We investigated this phenomenon and find that leak expression is mediated by both infrequent transcription from the inverted transgene in recombinant-dependent AAV designs and recombination events during bacterial AAV plasmid production...
December 16, 2019: Proceedings of the National Academy of Sciences of the United States of America
Shaghayegh Navabpour, Janine L Kwapis, Timothy J Jarome
The past decade has produced an explosion in the number and variety of genetic tools available to neuroscientists, resulting in an unprecedented ability to precisely manipulate the genome and epigenome in behaving animals. However, no single resource exists that describes all of the tools available to neuroscientists. Here, we review the genetic, transgenic, and viral techniques that are currently available to probe the complex relationship between genes and cognition. Topics covered include types of traditional transgenic mouse models (knockout, knock-in, reporter lines), inducible systems (Cre-loxP, Tet-On, Tet-Off) and cell- and circuit-specific systems (TetTag, TRAP, DIO-DREADD)...
December 13, 2019: Neuroscience and Biobehavioral Reviews
Chonprakun Thagun, Jo-Ann Chuah, Keiji Numata
The plastid is an organelle that functions as a cell factory to supply food and oxygen to the plant cell and is therefore a potential target for genetic engineering to acquire plants with novel photosynthetic traits or the ability to produce valuable biomolecules. Conventional plastid genome engineering technologies are laborious for the preparation of plant material, require expensive experimental instruments, and are time consuming for obtaining a transplastomic plant line that produces significant levels of the biomolecule of interest...
December 2019: Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
Baljeet Singh, Neha Salaria, Kajal Thakur, Sarvjeet Kukreja, Shristy Gautam, Umesh Goutam
Heat stress as a yield limiting issue has become a major threat for food security as global warming progresses. Being sessile, plants cannot avoid heat stress. They respond to heat stress by activating complex molecular networks, such as signal transduction, metabolite production and expressions of heat stress-associated genes. Some plants have developed an intricate signalling network to respond and adapt it. Heat stress tolerance is a polygenic trait, which is regulated by various genes, transcriptional factors, proteins and hormones...
2019: F1000Research
Anand K Singh, Akilu Abdullahi, Matthias Soller, Alexandre David, Saverio Brogna
The distribution of assembled, and potentially translating, ribosomes within cells can be visualised in Drosophila by using Bimolecular Fluorescence Complementation (BiFC) to monitor the interaction between tagged pairs of 40S and 60S ribosomal proteins (RPs) that are close neighbours across inter-subunit junctions in the assembled 80S ribosome. Here we describe transgenes expressing two novel RP pairs tagged with Venus-based BiFC fragments that considerably increase the sensitivity of this technique we termed Ribo-BiFC...
December 10, 2019: Biology Open
Harry O Orlans, Alun R Barnard, Robert E MacLaren
Imaging techniques have revolutionised the assessment of retinal disease in humans and animal models. Here we describe a novel technique for the in vivo visualisation of rod photoreceptors which permits semiquantitative assessment of outer retinal degeneration, and validate this approach in two mouse models of retinitis pigmentosa (RP). Transgenic mice carrying an Nrl-EGFP allele and homozygous for either knock-out of rhodopsin (Nrl-EGFP, Rho-/- ) or heterozygous for knock-in of P23H mutant rhodopsin (Nrl-EGFP, RhoP23H/+ ) were used in this study...
December 6, 2019: Experimental Eye Research
Hassan M M Ahmed, Luisa Hildebrand, Ernst A Wimmer
BACKGROUND: The invasive fruit pest Drosophila suzukii was reported for the first time in Europe and the USA in 2008 and has spread since then. The adoption of type II clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) as a tool for genome manipulation provides new ways to develop novel biotechnologically-based pest control approaches. Stage or tissue-specifically expressed genes are of particular importance in the field of insect biotechnology...
December 5, 2019: BMC Biotechnology
Ajantha Sinniah, Samia Yazid, Stefania Bena, Sonia M Oliani, Mauro Perretti, Rod J Flower
Mast cell stabilizers like cromoglycate and nedocromil are mainstream treatments for ocular allergy. Biochemical studies in vitro suggest that these drugs prevent mast cell degranulation through the release of Annexin-A1 (Anx-A1) protein. However, the direct effect of Anx-A1 gene deletion on mast cell function in vitro and in vivo is yet to be fully investigated. Hence, we aim to elucidate the role of Anx-A1 in mast cell function, both in vivo and in vitro , using a transgenic mouse model where the Anx-A1 gene has been deleted...
2019: Frontiers in Pharmacology
Borys Frankewycz, Daniel Cimino, Nelly Andarawis-Puri
BACKGROUND: Tendon injuries are common musculoskeletal injuries that heal with scar tissue formation, often achieving reduced biomechanical and functional properties. The murine patellar tendon is a research tool that holds potential for investigating tendon healing and can be useful for exploring therapeutic strategies. Since healing is a complex process that results from the collaboration between the systemic and local tissue environment, a murine tendon transplantation model that can be applied to transgenic mice and genetic mutants would allow isolation of systemic versus local tendon factors in driving effective tendon healing...
December 2, 2019: Journal of Orthopaedic Surgery and Research
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