22 research outputs found
One-step Multiplex Transgenesis via Sleeping Beauty Transposition in Cattle
Genetically modified cattle are important for developing new biomedical models and for an improved understanding of the pathophysiology of zoonotic diseases. However, genome editing and genetic engineering based on somatic cell nuclear transfer suffer from a low overall efficiency. Here, we established a highly efficient one-step multiplex gene transfer system into the bovine genome.Fil: Garrels, Wiebke. Institut für Nutztiergenetik; AlemaniaFil: Talluri, Thirumala R.. Institut für Nutztiergenetik; AlemaniaFil: Apfelbaum, Ronja. Institut für Nutztiergenetik; AlemaniaFil: Carratalá, Yanet P.. Institut für Nutztiergenetik; AlemaniaFil: Bosch, Pablo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Pötzsch, Kerstin. Paul Ehrlich Institute; AlemaniaFil: Grueso, Esther. Paul Ehrlich Institute; AlemaniaFil: Ivics, Zoltan. Paul Ehrlich Institute; AlemaniaFil: Kues, Wilfred. Institut für Nutztiergenetik; Alemani
Some reproductive parameters of Zanskari mares reared in an organized farm under tropical climate
The present study was undertaken to record the data on duration of estrus, estrus cycle length, size of the follicle at ovulation, period of gestation and estrus after foaling (foal heat) in respect of Zanskari mares for 4 consecutive breeding seasons (2010–2014). The mares were closely monitored with transrectal ultrasonography for their follicle size variation and to determine size of the follicle at ovulation. The mares were inseminated with frozen thawed semen from Zanskari stallions. This is the first time ever that Zanskari stallion semen was cryopreserved successfully and viable foals were produced by using frozen semen via artificial insemination. The reproductive parameters of the mares of Zanskari breed in India are poorly investigated and thus no reports are available on reproductive characteristics of Zanskari breed till today in the literature. Our results can be useful as a ready reference for some of the reproductive characteristics and traits of Zanskari breed as well as indigenous horses reared under tropical region. The reproductive parameters analysed in the present study are fitting within the characteristics of the other Indian horse breeds reared under this region and the differences may be caused by breed effect and other environmental factors
Establishment of cell-based transposon-mediated transgenesis in cattle
Transposon-mediated transgenesis is a well-established tool for genome modification in small animal models. However, translation of this active transgenic method to large animals warrants further investigations. Here, the piggyBac (PB) and sleeping beauty (SB) transposon systems were assessed for stable gene transfer into the cattle genome. Bovine fibroblasts were transfected either with a helper-independent PB system or a binary SB system. Both transposons were highly active in bovine cells increasing the efficiency of DNA integration up to 88 times over basal nonfacilitated integrations in a colony formation assay. SB transposase catalyzed multiplex transgene integrations in fibroblast cells transfected with the helper vector and two donor vectors carrying different transgenes (fluorophore and neomycin resistance). Stably transfected fibroblasts were used for SCNT and on in vitro embryo culture, morphologically normal blastocysts that expressed the fluorophore were obtained with both transposon systems. The data indicate that transpositionis a feasible approach for genetic engineering in the cattle genome.Fil: Alessio, Ana Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Fili, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Garrels, Wiebke. Institut für Nutztiergenetik; Alemania. Gottfried Wilhelm Leibniz Universität Hannover; AlemaniaFil: Forcato, Diego Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Olmos Nicotra, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Liaudat, Ana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Savy, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Hiriart, María Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Talluri, Thirumala R.. Institut für Nutztiergenetik; AlemaniaFil: Owens, Jesse B.. University of Hawaii at Manoa; Estados UnidosFil: Ivics, Zoltán. Paul-Ehrlich-Institute; AlemaniaFil: Salamone, Daniel Felipe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Moisyadi, Stefan. University of Hawaii at Manoa; Estados UnidosFil: Kues, Wilfried A.. Institut für Nutztiergenetik; AlemaniaFil: Bosch, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentin
Transposon-based reprogramming to induced pluripotency
Induced pluripotent stem (iPS) cells
represent a recent innovation in the field of stem cells.
Commonly, iPS cells are generated by viral transduction
of core reprogramming genes, such as Oct4, Sox2, Klf4,
c-Myc, Nanog and Lin28. However, integrating viruses,
like retro- and lentiviral vectors, may cause insertional
mutagenesis and may increase the risk of tumor
formation. Therefore, alternative methods which avoid
these safety concerns are intensively investigated. Here,
we review the current status of transposon-based
methods to induce pluripotency. DNA transposons are
non-viral elements, which can be effectively integrated
into a genome by their corresponding transposase
enzyme. The advantages of transposon-based gene
transfer are their increased safety, their large cargo
capacity, their relatively simple design, and the
availability of hyper-active and mutated transposase
enzymes. For example, integration-deficient, excisioncompetent transposase variants allow the complete
removal of the reprogramming transposon after
successful reprogramming to obtain transposon-free
reprogrammed cells. Transposon-based reprogramming
broaden the toolbox for iPS cell production and will
advance the establishment of safe, non-viral methods
Not Available
Not AvailableInduced pluripotent stem (iPS) cells
represent a recent innovation in the field of stem cells.
Commonly, iPS cells are generated by viral transduction
of core reprogramming genes, such as Oct4, Sox2, Klf4,
c-Myc, Nanog and Lin28. However, integrating viruses,
like retro- and lentiviral vectors, may cause insertional
mutagenesis and may increase the risk of tumor
formation. Therefore, alternative methods which avoid
these safety concerns are intensively investigated. Here,
we review the current status of transposon-based
methods to induce pluripotency. DNA transposons are
non-viral elements, which can be effectively integrated
into a genome by their corresponding transposase
enzyme. The advantages of transposon-based gene
transfer are their increased safety, their large cargo
capacity, their relatively simple design, and the
availability of hyper-active and mutated transposase
enzymes. For example, integration-deficient, excisioncompetent
transposase variants allow the complete
removal of the reprogramming transposon after
successful reprogramming to obtain transposon-free
reprogrammed cells. Transposon-based reprogramming
broaden the toolbox for iPS cell production and will
advance the establishment of safe, non-viral methods.Not Availabl
Not Available
Not AvailableInduced pluripotent stem (iPS) cells
represent a recent innovation in the field of stem cells.
Commonly, iPS cells are generated by viral transduction
of core reprogramming genes, such as Oct4, Sox2, Klf4,
c-Myc, Nanog and Lin28. However, integrating viruses,
like retro- and lentiviral vectors, may cause insertional
mutagenesis and may increase the risk of tumor
formation. Therefore, alternative methods which avoid
these safety concerns are intensively investigated. Here,
we review the current status of transposon-based
methods to induce pluripotency. DNA transposons are
non-viral elements, which can be effectively integrated
into a genome by their corresponding transposase
enzyme. The advantages of transposon-based gene
transfer are their increased safety, their large cargo
capacity, their relatively simple design, and the
availability of hyper-active and mutated transposase
enzymes. For example, integration-deficient, excisioncompetent
transposase variants allow the complete
removal of the reprogramming transposon after
successful reprogramming to obtain transposon-free
reprogrammed cells. Transposon-based reprogramming
broaden the toolbox for iPS cell production and will
advance the establishment of safe, non-viral methods.ICAR, DBT, NIA
Not Available
Not AvailableSemen ejaculates (8) were obtained from 2 Marwari stallions, 2 Zanskari stallions and 2 exotic jacks (martina
franca). Gel portion of the semen was removed by filtration and immediately the seminal plasma was separated by
centrifugation at 2000 rpm for 30 min and stored at –20°C until further analysis. Total protein from the seminal plasma
was estimated by Lowry method and the molecular weight was analysed by SDS-PAGE. The total protein concentration
was found to be 0.71, 2.24 and 2.21 g/dl in Marwari, Zanskari and exotic jack seminal plasma respectively. The differences
in protein concentration were found to be significantly higher in Zanskari and jack stallions as compared to Marwari
stallion. There was no significant difference of total protein values between Zanskari and jack stallions. A total of 12
protein bands were observed in the seminal plasma of both Marwari and Zanskari stallions and are in common, whereas,
15 protein bands were observed in the seminal plasma of exotic jacks. The seminal plasma proteins in Marwari and
Zanskari stallions ranged from 11.45KDa to 130.23kDa and that of exotic jack ranged from 11.45kDa to 162.83kDa.
Out of 15 protein bands observed, 3 were unique in jack stallions having molecular weight of 36.52 kDa, 39.25 kDa and
1602.83 kDa. It can be concluded from this study that at least 12 bands of protein were in common between the all the
3 breeds of equines, and seminal plasma protein profile of jack stallion is different from the Marwari and Zanskari
stallions.ICA