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Efficient Gene Transfer into Early Chicken Embryos by Electroporation of Stage X Blastoderms in Vivo, Applying Electric Pulses Vertically to the Blastoderm Layer

  In order to introduce exogenous DNA into early chicken embryos by transfecting the stage X (EG&K) blastoderm, in vivo electroporation was applied to it. A pair of novel electrodes was devised for applying electric pulses vertically to the blastoderm layer. Fertilised eggs at stage X were broken an... Full description

Journal Title: The Journal of Poultry Science 2002, Vol.39(4), p.292
Main Author: Naito, Mitsuru
Other Authors: Sano, Akiko , Tagami, Takahiro , Harumi, Takashi , Matsubara, Yuko , Kuwana, Takashi
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 13467395 ; E-ISSN: 13490486
Link: http://search.proquest.com/docview/1439289965/?pq-origsite=primo
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title: Efficient Gene Transfer into Early Chicken Embryos by Electroporation of Stage X Blastoderms in Vivo, Applying Electric Pulses Vertically to the Blastoderm Layer
format: Article
creator:
  • Naito, Mitsuru
  • Sano, Akiko
  • Tagami, Takahiro
  • Harumi, Takashi
  • Matsubara, Yuko
  • Kuwana, Takashi
subjects:
  • Blastoderm
  • Chick Embryo
  • Electroporation
  • Embryo Culture
  • Gfp Gene
ispartof: The Journal of Poultry Science, 2002, Vol.39(4), p.292
description:   In order to introduce exogenous DNA into early chicken embryos by transfecting the stage X (EG&K) blastoderm, in vivo electroporation was applied to it. A pair of novel electrodes was devised for applying electric pulses vertically to the blastoderm layer. Fertilised eggs at stage X were broken and the blastoderm was placed on top of the yolk. One of the electrodes was placed on the blastoderm layer through the yolk membrane, and the other electrode was placed on the bottom of the yolk. DNA (GFP gene) was injected into the blastoderm, and electric square pulses were applied 5 times at 5-100V for loading periods of 50 msec per pulse at one-second intervals. The manipulated embryos were transferred into host eggshells and incubated for 4 days. The viability of the manipulated embryos was 67.0% (59/88) on day 3 of incubation. The expression pattern of GFP gene was mostly mosaic in the embryos, and the percentages of embryos that expressed the GFP gene were 97.7% (86/88), 93.2% (82/88) and 75.0% (66/88) on days 1, 2 and 3 of incubation. A high rate of GFP gene expression was observed in the embryonic and extra-embryonic tissues (54.2%, 32/59) and also in the extra-embryonic tissues only (27.1%, 16/59) on day 3 of incubation. The expression of the GFP gene throughout the embryonic tissues was also observed. These results suggest that the system for applying electric pulses vertically to the blastoderm layer is effective in introducing exogenous DNA into early chicken embryos.
language: eng
source:
identifier: ISSN: 13467395 ; E-ISSN: 13490486
fulltext: fulltext
issn:
  • 13467395
  • 1346-7395
  • 13490486
  • 1349-0486
url: Link


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titleEfficient Gene Transfer into Early Chicken Embryos by Electroporation of Stage X Blastoderms in Vivo, Applying Electric Pulses Vertically to the Blastoderm Layer
creatorNaito, Mitsuru ; Sano, Akiko ; Tagami, Takahiro ; Harumi, Takashi ; Matsubara, Yuko ; Kuwana, Takashi
ispartofThe Journal of Poultry Science, 2002, Vol.39(4), p.292
identifierISSN: 13467395 ; E-ISSN: 13490486
description  In order to introduce exogenous DNA into early chicken embryos by transfecting the stage X (EG&K) blastoderm, in vivo electroporation was applied to it. A pair of novel electrodes was devised for applying electric pulses vertically to the blastoderm layer. Fertilised eggs at stage X were broken and the blastoderm was placed on top of the yolk. One of the electrodes was placed on the blastoderm layer through the yolk membrane, and the other electrode was placed on the bottom of the yolk. DNA (GFP gene) was injected into the blastoderm, and electric square pulses were applied 5 times at 5-100V for loading periods of 50 msec per pulse at one-second intervals. The manipulated embryos were transferred into host eggshells and incubated for 4 days. The viability of the manipulated embryos was 67.0% (59/88) on day 3 of incubation. The expression pattern of GFP gene was mostly mosaic in the embryos, and the percentages of embryos that expressed the GFP gene were 97.7% (86/88), 93.2% (82/88) and 75.0% (66/88) on days 1, 2 and 3 of incubation. A high rate of GFP gene expression was observed in the embryonic and extra-embryonic tissues (54.2%, 32/59) and also in the extra-embryonic tissues only (27.1%, 16/59) on day 3 of incubation. The expression of the GFP gene throughout the embryonic tissues was also observed. These results suggest that the system for applying electric pulses vertically to the blastoderm layer is effective in introducing exogenous DNA into early chicken embryos.
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titleEfficient Gene Transfer into Early Chicken Embryos by Electroporation of Stage X Blastoderms in Vivo, Applying Electric Pulses Vertically to the Blastoderm Layer
description  In order to introduce exogenous DNA into early chicken embryos by transfecting the stage X (EG&K) blastoderm, in vivo electroporation was applied to it. A pair of novel electrodes was devised for applying electric pulses vertically to the blastoderm layer. Fertilised eggs at stage X were broken and the blastoderm was placed on top of the yolk. One of the electrodes was placed on the blastoderm layer through the yolk membrane, and the other electrode was placed on the bottom of the yolk. DNA (GFP gene) was injected into the blastoderm, and electric square pulses were applied 5 times at 5-100V for loading periods of 50 msec per pulse at one-second intervals. The manipulated embryos were transferred into host eggshells and incubated for 4 days. The viability of the manipulated embryos was 67.0% (59/88) on day 3 of incubation. The expression pattern of GFP gene was mostly mosaic in the embryos, and the percentages of embryos that expressed the GFP gene were 97.7% (86/88), 93.2% (82/88) and 75.0% (66/88) on days 1, 2 and 3 of incubation. A high rate of GFP gene expression was observed in the embryonic and extra-embryonic tissues (54.2%, 32/59) and also in the extra-embryonic tissues only (27.1%, 16/59) on day 3 of incubation. The expression of the GFP gene throughout the embryonic tissues was also observed. These results suggest that the system for applying electric pulses vertically to the blastoderm layer is effective in introducing exogenous DNA into early chicken embryos.
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abstract  In order to introduce exogenous DNA into early chicken embryos by transfecting the stage X (EG&K) blastoderm, in vivo electroporation was applied to it. A pair of novel electrodes was devised for applying electric pulses vertically to the blastoderm layer. Fertilised eggs at stage X were broken and the blastoderm was placed on top of the yolk. One of the electrodes was placed on the blastoderm layer through the yolk membrane, and the other electrode was placed on the bottom of the yolk. DNA (GFP gene) was injected into the blastoderm, and electric square pulses were applied 5 times at 5-100V for loading periods of 50 msec per pulse at one-second intervals. The manipulated embryos were transferred into host eggshells and incubated for 4 days. The viability of the manipulated embryos was 67.0% (59/88) on day 3 of incubation. The expression pattern of GFP gene was mostly mosaic in the embryos, and the percentages of embryos that expressed the GFP gene were 97.7% (86/88), 93.2% (82/88) and 75.0% (66/88) on days 1, 2 and 3 of incubation. A high rate of GFP gene expression was observed in the embryonic and extra-embryonic tissues (54.2%, 32/59) and also in the extra-embryonic tissues only (27.1%, 16/59) on day 3 of incubation. The expression of the GFP gene throughout the embryonic tissues was also observed. These results suggest that the system for applying electric pulses vertically to the blastoderm layer is effective in introducing exogenous DNA into early chicken embryos.
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