The Experts below are selected from a list of 191919 Experts worldwide ranked by ideXlab platform
Piet A Van Rijn - One of the best experts on this subject based on the ideXlab platform.
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requirements and comparative analysis of reverse genetics for bluetongue virus btv and african horse sickness virus ahsv
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus (Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames.
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Requirements and comparative analysis of reverse genetics for bluetongue virus (BTV) and African horse sickness virus (AHSV)
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus ( Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames. Results Plasmids containing full length cDNA of the 10 genome segments for T7 promoter-driven production of full length run-off RNA transcripts and expression plasmids with optimized open reading frames (ORFs) were used. BTV and AHSV were rescued using reverse genetics. The requirement of each expression plasmid and capping of RNA transcripts for reverse genetics were studied and compared for BTV and AHSV. BTV was recovered by Transfection of VP1 and NS2 expression plasmids followed by Transfection of a set of ten capped RNAs. VP3 expression plasmid was also required if uncapped RNAs were transfected. Recovery of AHSV required Transfection of VP1, VP3 and NS2 expression plasmids followed by Transfection of capped RNA transcripts. Plasmid-driven expression of VP4, 6 and 7 was also needed when uncapped RNA transcripts were used. Irrespective of capping of RNA transcripts, NS1 expression plasmid was not needed for recovery, although NS1 protein is essential for virus propagation. Improvement of reverse genetics for AHSV was clearly demonstrated by rescue of several mutants and reassortants that were not rescued with previous methods. Conclusions A limited number of expression plasmids is required for rescue of BTV or AHSV using reverse genetics, making the system much more versatile and generally applicable. Optimization of reverse genetics enlarge the possibilities to rescue virus mutants and reassortants, and will greatly benefit the control of these important diseases of livestock and companion animals.
Rene G P Van Gennip - One of the best experts on this subject based on the ideXlab platform.
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requirements and comparative analysis of reverse genetics for bluetongue virus btv and african horse sickness virus ahsv
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus (Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames.
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Requirements and comparative analysis of reverse genetics for bluetongue virus (BTV) and African horse sickness virus (AHSV)
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus ( Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames. Results Plasmids containing full length cDNA of the 10 genome segments for T7 promoter-driven production of full length run-off RNA transcripts and expression plasmids with optimized open reading frames (ORFs) were used. BTV and AHSV were rescued using reverse genetics. The requirement of each expression plasmid and capping of RNA transcripts for reverse genetics were studied and compared for BTV and AHSV. BTV was recovered by Transfection of VP1 and NS2 expression plasmids followed by Transfection of a set of ten capped RNAs. VP3 expression plasmid was also required if uncapped RNAs were transfected. Recovery of AHSV required Transfection of VP1, VP3 and NS2 expression plasmids followed by Transfection of capped RNA transcripts. Plasmid-driven expression of VP4, 6 and 7 was also needed when uncapped RNA transcripts were used. Irrespective of capping of RNA transcripts, NS1 expression plasmid was not needed for recovery, although NS1 protein is essential for virus propagation. Improvement of reverse genetics for AHSV was clearly demonstrated by rescue of several mutants and reassortants that were not rescued with previous methods. Conclusions A limited number of expression plasmids is required for rescue of BTV or AHSV using reverse genetics, making the system much more versatile and generally applicable. Optimization of reverse genetics enlarge the possibilities to rescue virus mutants and reassortants, and will greatly benefit the control of these important diseases of livestock and companion animals.
Ren-xi Zhuo - One of the best experts on this subject based on the ideXlab platform.
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calcium phosphate dna co precipitates encapsulated fast degrading polymer films for substrate mediated gene delivery
Journal of Biomedical Materials Research Part B, 2009Co-Authors: Qiao Zhang, Dong Zhao, Xian-zheng Zhang, Si-xue Cheng, Ren-xi ZhuoAbstract:Calcium-phosphate/deoxyribose nucleic acid (Ca-P/DNA) co-precipitates were deposited on or encapsulated in fast-degrading polymer films with surface erosion degradation mechanism to mediate cell Transfection. The polymer, cholic acid functionalized star poly(DL-lactide), was synthesized through the ring-opening polymerization of DL-lactide initiated by cholic acid. The releases of DNA from the Ca-P/DNA co-precipitates deposited film and the Ca-P/DNA co-precipitates encapsulated film were determined and compared. The in vitro gene Transfections of HEK293 cells, Hela cells, and NIH 3T3 cells showed that the expression of pGL3-Luc plasmid could be effectively mediated by the Ca-P/DNA co-precipitates deposited and encapsulated polymer films. In addition, the films did not exhibit any additional cytotoxicity to the cells during the Transfections, indicating that the fast-degrading polymer films have great potential in localized gene delivery.
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three dimensional fast degrading polymer films for delivery of calcium phosphate dna co precipitates in solid phase Transfection
Journal of Materials Chemistry, 2009Co-Authors: Xian-zheng Zhang, Si-xue Cheng, Ren-xi ZhuoAbstract:We prepared Ca-P/DNA co-precipitates deposited and encapsulated in polymer films for substrate-mediated gene delivery. The polymer films were composed of fast-degrading functionalized star poly(DL-lactide) CA-PDLLA and block copolymer poly(L-lactide)-poly(ethylene glycol)-poly(L-lactide) (PLLA-PEG-PLLA). The in vitrogene Transfections of pGL3-Luc and pEGFP-C1 plasmids in HEK293T cells mediated by different films were studied. The effect of presence of a water-soluble polymer, poly-α,β-[N-(2-hydroxyethyl)-L-aspartamide], on the Transfection activity of the film-mediated Transfection was studied. The gene expressions could be effectively mediated by the Ca-P/DNA co-precipitates deposited and encapsulated in polymer films. During the cellular Transfection, the fast-degrading films rapidly released Ca-P/DNA co-precipitates to mediate gene Transfection, and the degradation products did not show any negative effects on the gene expression. The Ca-P/DNA co-precipitates deposited and encapsulated in films have promising applications in substrate-mediated gene delivery.
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dendrimer dna complexes encapsulated in a water soluble polymer and supported on fast degrading star poly dl lactide for localized gene delivery
Journal of Controlled Release, 2007Co-Authors: Si-xue Cheng, Xian-zheng Zhang, Ren-xi ZhuoAbstract:Abstract Polyamidoamine (PAMAM) dendrimer/DNA complexes encapsulated in a water soluble polymer, poly-α,β-[N-(2-hydroxyethyl)- l -aspartamide], were supported on a cholic acid functionalized star poly( dl -lactide) film with a fast degradation rate to mediate localized gene delivery. The in vitro gene Transfections of two types of cells, HEK293 and NIH3T3, were investigated. The expressions of pGL3-Luc and pEGFP-C1 plasmids in HEK293 cells indicated that the star poly( dl -lactide) supported PHEA encapsulated PAMAM/DNA complexes could effectively mediate Transfection, with Transfection efficiencies which were comparable to that of solution-based Transfections. Whereas the PAMAM/DNA complexes directly supported on the star poly( dl -lactide) film showed a much lower expression level for HEK293, which indicated the existence of PHEA played an important role in the efficient Transfection. The solid support-based Transfection for NIH3T3 cells exhibited higher expressions of pGL3-Luc compared with the solution-based Transfection. Encapsulating PAMAM/DNA complexes in PHEA could further improve the gene expression in NIH3T3. During the cellular Transfection, the degradation of the cholic acid functionalized star poly( dl -lactide) film could be obviously detected and the degradation did not show any unfavorable effects on the gene expression, which implied this solid support-based gene delivery device had great potential for localized Transfection.
Femke Feenstra - One of the best experts on this subject based on the ideXlab platform.
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requirements and comparative analysis of reverse genetics for bluetongue virus btv and african horse sickness virus ahsv
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus (Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames.
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Requirements and comparative analysis of reverse genetics for bluetongue virus (BTV) and African horse sickness virus (AHSV)
Virology Journal, 2016Co-Authors: Piet A Van Rijn, Sandra G P Van De Water, Femke Feenstra, Rene G P Van GennipAbstract:Background Bluetongue virus (BTV) and African horse sickness virus (AHSV) are distinct arthropod borne virus species in the genus Orbivirus ( Reoviridae family), causing the notifiable diseases Bluetongue and African horse sickness of ruminants and equids, respectively. Reverse genetics systems for these orbiviruses with their ten-segmented genome of double stranded RNA have been developed. Initially, two subsequent Transfections of in vitro synthesized capped run-off RNA transcripts resulted in the recovery of BTV. Reverse genetics has been improved by Transfection of expression plasmids followed by Transfection of ten RNA transcripts. Recovery of AHSV was further improved by use of expression plasmids containing optimized open reading frames. Results Plasmids containing full length cDNA of the 10 genome segments for T7 promoter-driven production of full length run-off RNA transcripts and expression plasmids with optimized open reading frames (ORFs) were used. BTV and AHSV were rescued using reverse genetics. The requirement of each expression plasmid and capping of RNA transcripts for reverse genetics were studied and compared for BTV and AHSV. BTV was recovered by Transfection of VP1 and NS2 expression plasmids followed by Transfection of a set of ten capped RNAs. VP3 expression plasmid was also required if uncapped RNAs were transfected. Recovery of AHSV required Transfection of VP1, VP3 and NS2 expression plasmids followed by Transfection of capped RNA transcripts. Plasmid-driven expression of VP4, 6 and 7 was also needed when uncapped RNA transcripts were used. Irrespective of capping of RNA transcripts, NS1 expression plasmid was not needed for recovery, although NS1 protein is essential for virus propagation. Improvement of reverse genetics for AHSV was clearly demonstrated by rescue of several mutants and reassortants that were not rescued with previous methods. Conclusions A limited number of expression plasmids is required for rescue of BTV or AHSV using reverse genetics, making the system much more versatile and generally applicable. Optimization of reverse genetics enlarge the possibilities to rescue virus mutants and reassortants, and will greatly benefit the control of these important diseases of livestock and companion animals.
Idoia Gallego - One of the best experts on this subject based on the ideXlab platform.
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non viral vectors based on cationic niosomes and minicircle dna technology enhance gene delivery efficiency for biomedical applications in retinal disorders
Nanomedicine: Nanotechnology Biology and Medicine, 2019Co-Authors: Idoia Gallego, Ilia Villatebeitia, Gema Martineznavarrete, Margarita Menendez, Tania Lopezmendez, Cristina Sotosanchez, Jon ZarateAbstract:Abstract Low Transfection efficiency is a major challenge to overcome in non-viral approaches to reach clinical practice. Our aim was to explore new strategies to achieve more efficient non-viral gene therapies for clinical applications and in particular, for retinal diseases. Cationic niosomes and three GFP-encoding genetic materials consisting on minicircle (2.3 kb), its parental plasmid (3.5 kb) and a larger plasmid (5.5 kb) were combined to form nioplexes. Once fully physicochemically characterized, in vitro experiments in ARPE-19 retina epithelial cells showed that Transfection efficiency of minicircle nioplexes doubled that of plasmids ones, maintaining good cell viability in all cases. Transfections in retinal primary cells and injections of nioplexes in rat retinas confirmed the higher capacity of cationic niosomes vectoring minicircle to deliver the genetic material into retina cells. Therefore, nioplexes based on cationic niosomes vectoring minicircle DNA represent a potential tool for the treatment of inherited retinal diseases.
