High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model
This study reports the high efficiencies of gene delivery and gene targeting (GT) in a medaka fish embryonic stem (ES) cell model. A baculoviral (BV) vector is demonstrated to significantly improve gene delivery in medaka ES (MES I) cells, Furthermore, the BV mediated host integration does not compr...
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sg-ntu-dr.10356-465302023-03-03T16:05:19Z High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model Yan, Yan Hong YunHan Li Changming School of Chemical and Biomedical Engineering Centre for Advanced Bionanosystems DRNTU::Engineering::Chemical engineering::Biotechnology This study reports the high efficiencies of gene delivery and gene targeting (GT) in a medaka fish embryonic stem (ES) cell model. A baculoviral (BV) vector is demonstrated to significantly improve gene delivery in medaka ES (MES I) cells, Furthermore, the BV mediated host integration does not compromise genome stability and pluripotency of transduced cells. GT via homologous recombination (HR) can generate precise genetic alterations in vertebrate genomes and has long been limited in mouse ES cells. It remains unresolved whether GT can also be developed in lower vertebrates. By using p53 as a gene model, the correct HR events are revealed by PCR, Southern, FISH and sequencing analyses, and a 22.8% GT efficiency is obtained in MES I cells. The pluripotency of targeted ES cells are well retained. These results demonstrate that highly efficient GT can be reproducibly obtained in MES I cells, offering a lower vertebrate model organism for GT towards knock-out animal production. DOCTOR OF PHILOSOPHY (SCBE) 2011-12-13T08:07:17Z 2011-12-13T08:07:17Z 2011 2011 Thesis Yan, Y. (2011). High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/46530 10.32657/10356/46530 en 166 p. application/pdf |
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DRNTU::Engineering::Chemical engineering::Biotechnology Yan, Yan High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
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This study reports the high efficiencies of gene delivery and gene targeting (GT) in a medaka fish embryonic stem (ES) cell model. A baculoviral (BV) vector is demonstrated to significantly improve gene delivery in medaka ES (MES I) cells, Furthermore, the BV mediated host integration does not compromise genome stability and pluripotency of transduced cells. GT via homologous recombination (HR) can generate precise genetic alterations in vertebrate genomes and has long been limited in mouse ES cells. It remains unresolved whether GT can also be developed in lower vertebrates. By using p53 as a gene model, the correct HR events are revealed by PCR, Southern, FISH and sequencing analyses, and a 22.8% GT efficiency is obtained in MES I cells. The pluripotency of targeted ES cells are well retained. These results demonstrate that highly efficient GT can be reproducibly obtained in MES I cells, offering a lower vertebrate model organism for GT towards knock-out animal production. |
author2 |
Hong YunHan |
author_facet |
Hong YunHan Yan, Yan |
format |
Theses and Dissertations |
author |
Yan, Yan |
author_sort |
Yan, Yan |
title |
High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
title_short |
High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
title_full |
High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
title_fullStr |
High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
title_full_unstemmed |
High efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
title_sort |
high efficiencies of gene delivery and gene targeting in embryonic stem cells of the fish medaka as a lower vertebrate model |
publishDate |
2011 |
url |
https://hdl.handle.net/10356/46530 |
_version_ |
1759856708174217216 |