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Transgenic Research

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https://www.readbyqxmd.com/read/27905063/efficient-gene-targeting-in-mouse-zygotes-mediated-by-crispr-cas9-protein
#1
Chris J Jung, Junli Zhang, Elizabeth Trenchard, Kent C Lloyd, David B West, Barry Rosen, Pieter J de Jong
The CRISPR/Cas9 system has rapidly advanced targeted genome editing technologies. However, its efficiency in targeting with constructs in mouse zygotes via homology directed repair (HDR) remains low. Here, we systematically explored optimal parameters for targeting constructs in mouse zygotes via HDR using mouse embryonic stem cells as a model system. We characterized several parameters, including single guide RNA cleavage activity and the length and symmetry of homology arms in the construct, and we compared the targeting efficiency between Cas9, Cas9nickase, and dCas9-FokI...
November 30, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27900537/rnai-mediated-resistance-to-rice-black-streaked-dwarf-virus-in-transgenic-rice
#2
Mohamed M S Ahmed, Shiquan Bian, Muyue Wang, Jing Zhao, Bingwei Zhang, Qiaoquan Liu, Changquan Zhang, Shuzhu Tang, Minghong Gu, Hengxiu Yu
Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus in the family Reoviridae, causes significant economic losses in rice production in China and many other Asian countries. Development of resistant varieties by using conventional breeding methods is limited, as germplasm with high level of resistance to RBSDV have not yet been found. One of the most promising methods to confer resistance against RBSDV is the use of RNA interference (RNAi) technology. RBSDV non-structural protein P7-2, encoded by S7-2 gene, is a potential F-box protein and involved in the plant-virus interaction through the ubiquitination pathway...
November 30, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27900536/t%C3%AE-4-overexpression-based-on-the-piggybac-transposon-system-in-cashmere-goats-alters-hair-fiber-characteristics
#3
Bingbo Shi, Qiang Ding, Xiaolin He, Haijing Zhu, Yiyuan Niu, Bei Cai, Jiao Cai, Anming Lei, Danju Kang, Hailong Yan, Baohua Ma, Xiaolong Wang, Lei Qu, Yulin Chen
Increasing cashmere yield is one of the vital aims of cashmere goats breeding. Compared to traditional breeding methods, transgenic technology is more efficient and the piggyBac (PB) transposon system has been widely applied to generate transgenic animals. For the present study, donor fibroblasts were stably transfected via a PB donor vector containing the coding sequence of cashmere goat thymosin beta-4 (Tβ4) and driven by a hair follicle-specific promoter, the keratin-associated protein 6.1 (KAP6.1) promoter...
November 29, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27896535/generation-of-gene-edited-birds-in-one-generation-using-sperm-transfection-assisted-gene-editing-stage
#4
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)...
November 28, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27891561/citrus-psorosis-virus-coat-protein-derived-hairpin-construct-confers-stable-transgenic-resistance-in-citrus-against-psorosis-a-and-b-syndromes
#5
A De Francesco, N Costa, M L García
Citrus psorosis virus (CPsV) is the causal agent of psorosis, a serious and widespread citrus disease. Two syndromes of psorosis, PsA and PsB, have been described. PsB is the most aggressive and rampant form. Previously, we obtained Pineapple sweet orange plants transformed with a hairpin construct derived from the CPsV coat protein gene (ihpCP). Some of these plants were resistant to CPsV 90-1-1, a PsA isolate homologous to the transgene. In this study, we found that expression of the ihpCP transgene and siRNA production in lines ihpCP-10 and -15 were stable with time and propagation...
November 28, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27888434/a-novel-heavy-metal-atpase-peptide-from-prosopis-juliflora-is-involved-in-metal-uptake-in-yeast-and-tobacco
#6
Nisha S Keeran, G Ganesan, Ajay K Parida
Heavy metal pollution of agricultural soils is one of the most severe ecological problems in the world. Prosopis juliflora, a phreatophytic tree species, grows well in heavy metal laden industrial sites and is known to accumulate heavy metals. Heavy Metal ATPases (HMAs) are ATP driven heavy metal pumps that translocate heavy metals across biological membranes thus helping the plant in heavy metal tolerance and phytoremediation. In the present study we have isolated and characterized a novel 28.9 kDa heavy metal ATPase peptide (PjHMT) from P...
November 25, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27873161/fgf-treatment-of-host-embryos-injected-with-es-cells-increases-rates-of-chimaerism
#7
Cathérine Dupont, Friedemann Loos, John Kong-A-San, Joost Gribnau
In spite of the emergence of genome editing tools, ES cell mediated transgenesis remains the most controllable way of creating genetically modified animals. Although tetraploid (4N) complementation of 4N host embryos and ES cells, is the only method guaranteeing that offspring are entirely ES cell derived, this technique is challenging, not always successful and difficult to implement in some laboratory settings. The current study shows that pretreatment of host blastocysts with FGF4 prior to ES cell injection can provide an alternative method for the generation of animals displaying high rates of chimaerism...
November 21, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27838898/generation-of-mouse-lines-with-conditionally-or-constitutively-inactivated-snca-gene-and-rosa26-stop-lacz-reporter-located-in-cis-on-the-mouse-chromosome-6
#8
Andrei Yu Roman, Galina Limorenko, Alexey A Ustyugov, Tatiana V Tarasova, Ekaterina A Lysikova, Vladimir L Buchman, Natalia Ninkina
α-Synuclein is involved in many important molecular processes in neuronal cells and their synapses, and its malfunction has been linked to the development of Parkinson's and certain other neurodegenerative diseases. Animal models allowing tightly monitored conditional inactivation of the encoding gene, Snca, are indispensible for studies aimed at understanding normal function of α-synuclein in various neuronal populations and its role in pathogenesis of neurodegenerative diseases. We have recently reported the production of several novel mouse lines for manipulating expression of the endogenous Snca gene, including a line for Cre-recombinase-driven conditional inactivation of the gene (mice with floxed Snca) and a new line with a constitutive knockout of α-synuclein...
November 12, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27832434/monitoring-of-venus-transgenic-cell-migration-during-pregnancy-in-non-transgenic-rabbits
#9
N Lipták, O I Hoffmann, A Kerekes, G Iski, D Ernszt, K Kvell, L Hiripi, Z Bősze
Cell transfer between mother and fetus were demonstrated previously in several species which possess haemochorial placenta (e.g. in humans, mice, rats, etc.). Here we report the assessment of fetal and maternal microchimerism in non-transgenic (non-TG) New Zealand white rabbits which were pregnant with transgenic (TG) fetuses and in non-TG newborns of TG does. The TG construct, including the Venus fluorophore cDNA driven by a ubiquitous cytomegalovirus enhancer, chicken ß-actin promoter (CAGGS), was previously integrated into the rabbit genome by Sleeping Beauty transposon system...
November 10, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27830476/production-of-heterozygous-alpha-1-3-galactosyltransferase-ggta1-knock-out-transgenic-miniature-pigs-expressing-human-cd39
#10
Kimyung Choi, Joohyun Shim, Nayoung Ko, Heejong Eom, Jiho Kim, Jeong-Woong Lee, Dong-Il Jin, Hyunil Kim
Production of transgenic pigs for use as xenotransplant donors is a solution to the severe shortage of human organs for transplantation. The first barrier to successful xenotransplantation is hyperacute rejection, a rapid, massive humoral immune response directed against the pig carbohydrate GGTA1 epitope. Platelet activation, adherence, and clumping, all major features of thrombotic microangiopathy, are inevitable results of immune-mediated transplant rejection. Human CD39 rapidly hydrolyzes ATP and ADP to AMP; AMP is hydrolyzed by ecto-5'-nucleotidase (CD73) to adenosine, an anti-thrombotic and cardiovascular protective mediator...
November 9, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27807665/a-new-transgenic-mouse-model-for-conditional-overexpression-of-the-polycomb-group-protein-ezh2
#11
Martijn A J Koppens, Ellen Tanger, Karim Nacerddine, Bart Westerman, Ji-Ying Song, Maarten van Lohuizen
The Polycomb Group protein EZH2 is upregulated in most prostate cancers, and its overexpression is associated with poor prognosis. Most insights into the functional role of EZH2 in prostate cancer have been gained using cell lines and EZH2 inactivation studies. However, the question remains whether overexpression of EZH2 can initiate prostate tumourigenesis or drive tumour progression. Appropriate transgenic mouse models that are required to answer such questions are lacking. We developed one such transgenic mouse model for conditional overexpression of Ezh2...
November 2, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27787698/the-transgenic-expression-of-human-follistatin-344-increases-skeletal-muscle-mass-in-pigs
#12
Fei Chang, Rui Fang, Meng Wang, Xin Zhao, Wen Chang, Zaihu Zhang, Ning Li, Qingyong Meng
Follistatin (FST), which was first found in the follicles of cattle and pigs, has been shown to be an essential regulator for muscle development. Mice that were genetically engineered to overexpress Fst specifically in muscle had at least twice the amount of skeletal muscle mass as controls; these findings are similar to earlier results obtained in myostatin-knockout mice. However, the role of follistatin in skeletal muscle development has yet to be clarified in livestock. Here, we describe transgenic Duroc pigs that exogenously express Fst specifically in muscle tissue...
October 27, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27778153/phenotypic-characterization-of-a-novel-ho-1-depletion-model-in-the-rat
#13
Vassilios Atsaves, Maria G Detsika, Elpida Poulaki, Hara Gakiopoulou, Elias A Lianos
Although the protective role of HO-1 induction in various forms of kidney disease is well established, mechanisms other than heme catabolism to biliverdin, bilirubin and carbon monoxide have recently been identified. Unraveling these mechanisms requires the generation of appropriate animal models. The present study describes the generation of a HO-1 deficient Hmox1 (-/-) rat model and characterizes its renal and extrarenal phenotype. Hmox1 (-/-) rats had growth retardation and splenomegaly compared to their Hmox1 (+/+) littermates...
October 24, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27771868/proteome-rebalancing-in-transgenic-camelina-occurs-within-the-enlarged-proteome-induced-by-%C3%AE-carotene-accumulation-and-storage-protein-suppression
#14
Monica A Schmidt, Ken Pendarvis
Oilseed crops are global commodities for their oil and protein seed content. We have engineered the oilseed Camelina sativa to exhibit increased protein content with a slight decrease in oil content. The introduction of a phytoene synthase gene with an RNAi cassette directed to suppress the storage protein 2S albumin resulted in seeds with an 11-24 % elevation in overall protein. The phytoene synthase cassette alone produced enhanced β-carotene content of an average 275 ± 6.10 μg/g dry seed and an overall altered seed composition of 11 % less protein and comparable nontransgenic amounts of both oil and carbohydrates...
October 22, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27771867/plant-characterization-of-genetically-modified-maize-hybrids-mon-89%C3%A3-34-3%C3%A2-%C3%A3-%C3%A2-mon-88%C3%A3-17-3-mon-89%C3%A3-34-3%C3%A2-%C3%A3-%C3%A2-mon-%C3%A3-%C3%A3-6%C3%A3-3-6-and-mon-%C3%A3-%C3%A3-6%C3%A3-3-6-alternatives-for-maize-production-in-mexico
#15
Oscar Heredia Díaz, José Luis Aldaba Meza, Baltazar M Baltazar, Germán Bojórquez Bojórquez, Luciano Castro Espinoza, José Luis Corrales Madrid, Juan Manuel de la Fuente Martínez, Héctor Abel Durán Pompa, José Alonso Escobedo, Armando Espinoza Banda, José Antonio Garzón Tiznado, Juvencio González García, José Luis Guzmán Rodríguez, Jesús Ignacio Madueño Martínez, José Luis Martínez Carrillo, Chen Meng, Francisco Javier Quiñones Pando, Enrique Rosales Robles, Ignacio Ruiz Hernández, José Elías Treviño Ramírez, Hugo Raúl Uribe Montes, Francisco Zavala García
Environmental risk assessment (ERA) of genetically modified (GM) crops is a process to evaluate whether the biotechnology trait(s) in a GM crop may result in increased pest potential or harm to the environment. In this analysis, two GM insect-resistant (IR) herbicide-tolerant maize hybrids (MON-89Ø34-3 × MON-88Ø17-3 and MON-89Ø34-3 × MON-ØØ6Ø3-6) and one herbicide-tolerant GM hybrid (MON-ØØ6Ø3-6) were compared with conventional maize hybrids of similar genetic backgrounds. Two sets of studies, Experimental Phase and Pilot Phase, were conducted across five ecological regions (ecoregions) in Mexico during 2009-2013, and data were subject to meta-analysis...
October 22, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27744533/zygote-injection-of-crispr-cas9-rna-successfully-modifies-the-target-gene-without-delaying-blastocyst-development-or-altering-the-sex-ratio-in-pigs
#16
Kristin M Whitworth, Joshua A Benne, Lee D Spate, Stephanie L Murphy, Melissa S Samuel, Clifton N Murphy, Jürgen A Richt, Eric Walters, Randall S Prather, Kevin D Wells
The CRISPR/Cas9 genome editing tool has increased the efficiency of creating genetically modified pigs for use as biomedical or agricultural models. The objectives were to determine if DNA editing resulted in a delay in development to the blastocyst stage or in a skewing of the sex ratio. Six DNA templates (gBlocks) that were designed to express guide RNAs that target the transmembrane protease, serine S1, member 2 (TMPRSS2) gene were in vitro transcribed. Pairs of CRISPR guide RNAs that flanked the start codon and polyadenylated Cas9 were co-injected into the cytoplasm of zygotes and cultured in vitro to the blastocyst stage...
October 15, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27743222/reproductive-parameters-of-double-transgenic-zebrafish-danio-rerio-males-overexpressing-both-the-growth-hormone-gh-and-its-receptor-ghr
#17
Ana Cecilia Gomes Silva, Daniela Volcan Almeida, Bruna Felix Nornberg, Jessica Ribeiro Pereira, Diego Martins Pires, Carine Dahl Corcini, Antonio Sergio Varela Junior, Luis Fernando Marins
Growth hormone (GH) transgenesis presents a high potential application in aquaculture. However, excess GH may have serious consequences due to pleiotropic actions. In order to study these effects in zebrafish (Danio rerio), two transgenic lines were developed. The first expresses GH ubiquitously and constitutively (F0104 line), while the second expresses the GH receptor in a muscle-specific manner (Myo-GHR line). Results from the F0104 line showed accelerated growth but increased reproductive difficulties, while Myo-GHR did not show the expected increase in muscle mass...
October 14, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27730498/tamoxifen-dosing-for-cre-mediated-recombination-in-experimental-bronchopulmonary-dysplasia
#18
Jordi Ruiz-Camp, José Alberto Rodríguez-Castillo, Susanne Herold, Konstantin Mayer, István Vadász, Michelle D Tallquist, Werner Seeger, Katrin Ahlbrecht, Rory E Morty
Bronchopulmonary dysplasia (BPD) is the most common complication of preterm birth characterized by blunted post-natal lung development. BPD can be modelled in mice by exposure of newborn mouse pups to elevated oxygen levels. Little is known about the mechanisms of perturbed lung development associated with BPD. The advent of transgenic mice, where genetic rearrangements can be induced in particular cell-types at particular time-points during organogenesis, have great potential to explore the pathogenic mechanisms at play during arrested lung development...
October 11, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27696149/generation-of-il10-and-tgfb1-coexpressed-mice-displaying-resistance-to-ovalbumin-induced-asthma
#19
Fei Xu, Xiuhua Kang, Liang Chen, Chuanhui Chen, Gen Hu, Wei Bai, Wei Zhang
Asthma is a common chronic inflammatory disease in the airways with wide prevalence, and it is thought to be caused by the combinational factors in environment and genetics. A large body of studies has suggested that cell immunity played a vital role in regulating the airway hyperreactivity (AHR) and inflammation. Therefore, we here developed a mouse model of asthma by microinjecting the pronucleus with a vector spontaneously coding human IL10 and TGFB1 gene to explore the possible interaction between these two potent molecules during asthma progression...
October 1, 2016: Transgenic Research
https://www.readbyqxmd.com/read/27687031/transgenic-expression-of-phytase-in-wheat-endosperm-increases-bioavailability-of-iron-and-zinc-in-grains
#20
Nabeela Abid, Asia Khatoon, Asma Maqbool, Muhammad Irfan, Aftab Bashir, Irsa Asif, Muhammad Shahid, Asma Saeed, Henrik Brinch-Pedersen, Kauser A Malik
Phytate is a major constituent of wheat seeds and chelates metal ions, thus reducing their bioavailability and so the nutritional value of grains. Transgenic plants expressing heterologous phytase are expected to enhance degradation of phytic acid stored in seeds and are proposed to increase the in vitro bioavailability of mineral nutrients. Wheat transgenic plants expressing Aspergillus japonicus phytase gene (phyA) in wheat endosperm were developed till T3 generation. The transgenic lines exhibited 18-99 % increase in phytase activity and 12-76 % reduction of phytic acid content in seeds...
September 29, 2016: Transgenic Research
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