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Transgenic Research

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https://www.readbyqxmd.com/read/28634819/establishment-of-a-tagged-variant-of-lgr4-receptor-suitable-for-functional-and-expression-studies-in-the-mouse
#1
Vitezslav Kriz, Michaela Krausova, Petra Buresova, Jan Dobes, Dusan Hrckulak, Olga Babosova, Jiri Svec, Vladimir Korinek
Leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) is produced in a broad spectrum of mouse embryonic and adult tissues and its deficiency results in embryonal or perinatal lethality. The LGR4 function was mainly related to its potentiation of canonical Wnt signaling; however, several recent studies associate LGR4 with additional signaling pathways. To obtain a suitable tool for studying the signaling properties of Lgr4, we generated a tagged variant of the Lgr4 receptor using gene targeting in the mouse oocyte...
June 20, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28631185/the-increased-expression-of-follicle-stimulating-hormone-leads-to-a-decrease-of-fecundity-in-transgenic-large-white-female-pigs
#2
Kai Jiang, Pan Xu, Wanbo Li, Qiang Yang, Longyun Li, Chuanmin Qiao, Huanfa Gong, Hao Zheng, Zhimin Zhou, Hao Fu, Qiuyan Li, Yuyun Xing, Jun Ren
Follicle-stimulating hormone (FSH) is a pituitary gonadotropin regulating reproduction in mammals. Overexpression of the exogenous FSHα/β genes from Chinese Erhualian pigs improved female fecundity of transgenic (TG) mice and male spermatogenesis ability of Large White TG boars. Here, we investigated the impact of the exogenous FSHα/β genes on female reproductive performance of Large White TG pigs. First, we identified the integration site of the exogenous FSHα/β genes at 140,646,456 bp on chromosome 9 in these TG pigs using whole-genome sequencing...
June 19, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28608322/long-evans-rat-spermatogonial-lines-are-effective-germline-vectors-for-transgenic-rat-production
#3
F Kent Hamra, Christopher T Richie, Brandon K Harvey
Long Evans rat strains are applied as research models in a broad spectrum of biomedical fields (>15,800 citations, NCBI PubMed). Here, we report an approach to genetically modify the Long Evans rat germline in donor spermatogonial stem cells. Long Evans rat spermatogonial lines were derived from freshly isolated laminin-binding spermatogonia. Laminin-binding spermatogonia were cultured over multiple passages on fibroblast feeder layers in serum-free culture medium containing GDNF and FGF2. Long Evans rat spermatogonial lines were genetically modified by transposon transduction to express a germline, tdTomato reporter gene...
June 12, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28553699/generation-of-cmahko-gtko-shtnfri-fc-ho-1-quadruple-gene-modified-pigs
#4
Geon A Kim, Eun Mi Lee, Jun-Xue Jin, Sanghoon Lee, Anukul Taweechaipaisankul, Jong Ik Hwang, Zahid Alam, Curie Ahn, Byeong Chun Lee
As an alternative source of organs for transplantation into humans, attention has been directed to pigs due to their similarities in biological features and organ size. However, severe immune rejection has prevented successful xenotransplantation using pig organs and tissues. To overcome immune rejection, recently developed genetic engineering systems such as TALEN coupled with somatic cell nuclear transfer (SCNT) to make embryos could be used to produce pigs compatible with xenotransplantation. We used the TALEN system to target the non-Gal antigen cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene in pigs that is naturally deleted in humans...
May 28, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28547343/msmir156-affects-global-gene-expression-and-promotes-root-regenerative-capacity-and-nitrogen-fixation-activity-in-alfalfa
#5
Banyar Aung, Ruimin Gao, Margaret Y Gruber, Ze-Chun Yuan, Mark Sumarah, Abdelali Hannoufa
MicroRNA156 (miR156) regulates a network of downstream genes to affect plant growth and development. We previously generated alfalfa (Medicago sativa) plants that overexpress homologous miR156 (MsmiR156OE), and identified three of its SPL target genes. These plants exhibited increased vegetative yield, delayed flowering and longer roots. In this study, we aimed to elucidate the effect of miR156 on the root system, including effect on nodulation and nitrogen fixation. We found that MsmiR156 overexpression increases root regeneration capacity in alfalfa, but with little effect on root biomass at the early stages of root development...
May 25, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28534229/production-of-a-gonadotropin-releasing-hormone-2-receptor-knockdown-gnrhr2-kd-swine-line
#6
A T Desaulniers, R A Cederberg, G A Mills, C A Lents, B R White
Swine are the only livestock species that produce both the second mammalian isoform of gonadotropin-releasing hormone (GNRH2) and its receptor (GNRHR2). Previously, we reported that GNRH2 and GNRHR2 mediate LH-independent testosterone secretion from porcine testes. To further explore this ligand-receptor complex, a pig model with reduced GNRHR2 expression was developed. Small hairpin RNA sequences targeting porcine GNRHR2 were subcloned into a lentiviral-based vector, lentiviral particles were generated and microinjected into the perivitelline space of zygotes, and embryos were transferred into a recipient...
May 22, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28526984/reduced-weed-seed-shattering-by-silencing-a-cultivated-rice-gene-strategic-mitigation-for-escaped-transgenes
#7
Huanxin Yan, Lei Li, Ping Liu, Xiaoqi Jiang, Lei Wang, Jia Fang, Zhimin Lin, Feng Wang, Jun Su, Bao-Rong Lu
Transgene flow form a genetically engineered (GE) crop to its wild relatives may result in unwanted environmental consequences. Mitigating transgenes via introducing a gene that is disadvantageous to wild relatives but beneficial to crops, and is tightly-linked with the target transgenes, may provide a promising solution to limit the spread of transgenes in wild/weedy populations. Here we demonstrate a novel system with significantly reduced seed shattering in crop-weed hybrid descendants by partially silenced expression of the seed-shattering gene SH4 in cultivated rice, using artificial microRNA and antisense RNA techniques...
May 19, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28493168/comparative-statistical-component-analysis-of-transgenic-cyanophycin-producing-potatoes-in-greenhouse-and-field-trials
#8
Kerstin Schmidt, Jörg Schmidtke, Yvonne Mast, Eva Waldvogel, Wolfgang Wohlleben, Friederike Klemke, Wolfgang Lockau, Tina Hausmann, Maja Hühns, Inge Broer
Potatoes are a promising system for industrial production of the biopolymer cyanophycin as a second compound in addition to starch. To assess the efficiency in the field, we analysed the stability of the system, specifically its sensitivity to environmental factors. Field and greenhouse trials with transgenic potatoes (two independent events) were carried out for three years. The influence of environmental factors was measured and target compounds in the transgenic plants (cyanophycin, amino acids) were analysed for differences to control plants...
May 10, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28466411/assessment-of-the-potential-for-gene-flow-from-transgenic-maize-zea-mays-l-to-eastern-gamagrass-tripsacum-dactyloides-l
#9
Moon-Sub Lee, Eric K Anderson, Duška Stojšin, Marc A McPherson, Baltazar Baltazar, Michael J Horak, Juan Manuel de la Fuente, Kunsheng Wu, James H Crowley, A Lane Rayburn, D K Lee
Eastern gamagrass (Tripsacum dactyloides L.) belongs to the same tribe of the Poaceae family as maize (Zea mays L.) and grows naturally in the same region where maize is commercially produced in the USA. Although no evidence exists of gene flow from maize to eastern gamagrass in nature, experimental crosses between the two species were produced using specific techniques. As part of environmental risk assessment, the possibility of transgene flow from maize to eastern gamagrass populations in nature was evaluated with the objectives: (1) to assess the seeds of eastern gamagrass populations naturally growing near commercial maize fields for the presence of a transgenic glyphosate-tolerance gene (cp4 epsps) that would indicate cross-pollination between the two species, and (2) to evaluate the possibility of interspecific hybridization between transgenic maize used as male parent and eastern gamagrass used as female parent...
May 2, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28432544/expression-of-cphb-and-cphe-type-cyanophycinases-in-cyanophycin-producing-tobacco-and-comparison-of-their-ability-to-degrade-cyanophycin-in-plant-and-plant-extracts
#10
Daniel Ponndorf, Inge Broer, Henrik Nausch
Increasing the arginine (Arg) content in plants used as feed or food is of interest, since the supplementation of food with conditionally essential Arg has been shown to have nutritional benefits. An increase was achieved by the expression of the Arg-rich bacterial storage component, cyanophycin (CGP), in the chloroplast of transgenic plants. CGP is stable in plants and its degradation into β-aspartic acid (Asp)-Arg dipeptides, is solely catalyzed by bacterial cyanophycinases (CGPase). Dipeptides can be absorbed by animals even more efficiently than free amino acids (Matthews and Adibi 1976; Wenzel et al...
April 21, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28432545/genome-edited-animals-learning-from-gm-crops
#11
Ann Bruce
Genome editing of livestock is poised to become commercial reality, yet questions remain as to appropriate regulation, potential impact on the industry sector and public acceptability of products. This paper looks at how genome editing of livestock has attempted to learn some of the lessons from commercialisation of GM crops, and takes a systemic approach to explore some of the complexity and ambiguity in incorporating genome edited animals in a food production system. Current applications of genome editing are considered, viewed from the perspective of past technological applications...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28417275/rna-silencing-in-the-life-cycle-of-soybean-multiple-restriction-systems-and-spatiotemporal-variation-associated-with-plant-architecture
#12
Ayumi Mori, Hiroshi Sato, Megumi Kasai, Tetsuya Yamada, Akira Kanazawa
The expression of transgenes introduced into a plant genome is sometimes suppressed by RNA silencing. Although local and systemic spread of RNA silencing have been studied, little is known about the mechanisms underlying spatial and temporal variation in transgene silencing between individual plants or between plants of different generations, which occurs seemingly stochastically. Here, we analyzed the occurrence, spread, and transmission of RNA silencing of the green fluorescent protein (GFP) gene over multiple generations of the progeny of a single soybean transformant...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28409408/spontaneous-recombinase-activity-of-cre-ert2-in-vivo
#13
Jasmin Kristianto, Michael G Johnson, Ryley K Zastrow, Abigail B Radcliff, Robert D Blank
Inducible Cre-ERT recombinase technology is widely used for gene targeting studies. The second generation of inducible Cre-ERT recombinase, hemizygous B6.129S-Tg(UBC-cre/ERT2)1Ejb/J (hereafter abbreviated as Cre-ERT2), a fusion of a mutated estrogen receptor and Cre recombinase, was engineered to be more efficient and specific than the original Cre-ERT. The putative mechanism of selective Cre-mediated recombination is Cre sequestration in the cytoplasm in the basal state with translocation to the nucleus only in the presence of tamoxifen...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28332009/n-glycosylation-of-an-igg-antibody-secreted-by-nicotiana-tabacum-by-2-cells-can-be-modulated-through-co-expression-of-human-%C3%AE-1-4-galactosyltransferase
#14
Catherine Navarre, Nicolas Smargiasso, Laurent Duvivier, Joseph Nader, Johann Far, Edwin De Pauw, Marc Boutry
Nicotiana tabacum BY-2 suspension cells have several advantages that make them suitable for the production of full-size monoclonal antibodies which can be purified directly from the culture medium. Carbohydrate characterization of an antibody (Lo-BM2) expressed in N. tabacum BY-2 cells showed that the purified Lo-BM2 displays N-glycan homogeneity with a high proportion (>70%) of the complex GnGnXF glycoform. The stable co-expression of a human β-1,4-galactosyltransferase targeted to different Golgi sub-compartments altered Lo-BM2N-glycosylation and resulted in the production of an antibody that exhibited either hybrid structures containing a low abundance of the plant epitopes (α-1,3-fucose and β-1,2-xylose), or a large amount of galactose-extended N-glycan structures...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28326506/influence-of-calcareous-soil-on-cry3bb1-expression-and-efficacy-in-the-field
#15
David S Wangila, Arnubio Valencia J, Haichuan Wang, Blair D Siegfried, Lance J Meinke
Greater than expected injury by western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) to Cry3Bb1 expressing maize hybrids (Zea mays L.) has been reported in southwestern Nebraska. Affected areas of some fields are often associated with high pH calcareous soils where maize growth is poor and iron chlorosis is common. As part of a comprehensive study to understand potential causes of unexpected injury, experiments were conducted during 2013 and 2014 to ascertain whether the calcareous soil conditions and associated poor maize growth negatively affect the expression of Cry3Bb1...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28314980/ex-post-assessment-of-genetically-modified-low-level-presence-in-canadian-flax
#16
Helen M Booker, Eric G Lamb, Stuart J Smyth
Canada is the world's largest producer and exporter of flaxseed. In 2009, DNA from deregistered genetically modified (GM) CDC Triffid was detected in a shipment of Canadian flaxseed exported to Europe, causing a large decrease in the amount of flax planted in Canada and a major shift in export markets. The flax industry in Canada undertook major changes to ensure the removal of transgenic flax from the supply chain. To demonstrate compliance, Canada adopted a protocol involving testing grain samples (post-harvest) using an RT-PCR test for the construct found in CDC Triffid...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28243835/a-lifetime-aging-study-of-human-cd19-transgenic-mice
#17
W Iverson, E Straley, S Oldham, J Rojko, S Turman, Y Wang
Mice transgenic for human CD19 have been an important animal model to help understand the role of this molecule in B lymphocyte function. Previously, no lifetime studies had been performed to understand the effects of this CD19 over expression on the survival or spontaneous pathology within the C57BL/6J background strain. We conducted a lifetime study with interim sacrifices to understand the transgenic effects on clinical signs, body weight, survival, and spontaneous pathology. Blood and urine samples were collected from select animals at various time points during the study for measurement of clinical pathology parameters and groups of animals were euthanized and examined at predetermined intervals...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28105543/a-simple-dna-recombination-screening-method-by-rt-pcr-as-an-alternative-to-southern-blot
#18
LETTER
Eliene Albers, Mauro Sbroggiò, Javier Martin-Gonzalez, Alexandra Avram, Stephanie Munk, Andres J Lopez-Contreras
The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific genomic locus, but limits the detection of its correct genomic integration by standard PCR methods. Genomic Long Range PCR (LR-PCR), using primers adjacent to the homology arms, has been used as an alternative to radioactive-based Southern blot screenings...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/28070852/functional-characterization-of-the-powdery-mildew-susceptibility-gene-smmlo1-in-eggplant-solanum-melongena-l
#19
Valentina Bracuto, Michela Appiano, Luigi Ricciardi, Deniz Göl, Richard G F Visser, Yuling Bai, Stefano Pavan
Eggplant (Solanum melongena L.) is one of the most important vegetables among the Solanaceae and can be a host to fungal species causing powdery mildew (PM) disease. Specific homologs of the plant Mildew Locus O (MLO) gene family are PM susceptibility factors, as their loss of function results in a recessive form of resistance known as mlo resistance. In a previous work, we isolated the eggplant MLO homolog SmMLO1. SmMLO1 is closely related to MLO susceptibility genes characterized in other plant species. However, it displays a peculiar non-synonymous substitution that leads to a T → M amino acid change at protein position 422, in correspondence of the MLO calmodulin-binding domain...
June 2017: Transgenic Research
https://www.readbyqxmd.com/read/27896535/generation-of-gene-edited-birds-in-one-generation-using-sperm-transfection-assisted-gene-editing-stage
#20
Caitlin A Cooper, Arjun Challagulla, Kristie A Jenkins, Terry G Wise, Terri E O'Neil, Kirsten R Morris, Mark L Tizard, Timothy J Doran
Generating transgenic and gene edited mammals involves in vitro manipulation of oocytes or single cell embryos. Due to the comparative inaccessibility of avian oocytes and single cell embryos, novel protocols have been developed to produce transgenic and gene edited birds. While these protocols are relatively efficient, they involve two generation intervals before reaching complete somatic and germline expressing transgenic or gene edited birds. Most of this work has been done with chickens, and many protocols require in vitro culturing of primordial germ cells (PGCs)...
June 2017: Transgenic Research
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