A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells.
PiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its pot...
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089396&type=printable |
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| author | Jin-Bon Hong Fu-Ju Chou Amy T Ku Hsiang-Hsuan Fan Tung-Lung Lee Yung-Hsin Huang Tsung-Lin Yang I-Chang Su I-Shing Yu Shu-Wha Lin Chung-Liang Chien Hong-Nerng Ho You-Tzung Chen |
| author_facet | Jin-Bon Hong Fu-Ju Chou Amy T Ku Hsiang-Hsuan Fan Tung-Lung Lee Yung-Hsin Huang Tsung-Lin Yang I-Chang Su I-Shing Yu Shu-Wha Lin Chung-Liang Chien Hong-Nerng Ho You-Tzung Chen |
| author_sort | Jin-Bon Hong |
| collection | DOAJ |
| description | PiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its potential uses in functional genomics and regenerative medicine. Efforts to increase its transposition efficiency in mammals were made through engineering the corresponding transposase (PBase) codon usage to enhance its expression level and through screening for mutant PBase variants with increased enzyme activity. To improve the safety for its potential use in regenerative medicine applications, site-specific transposition was achieved by using engineered zinc finger- and Gal4-fused PBases. An excision-prone PBase variant has also been successfully developed. Here we describe the construction of a nucleolus-predominant PBase, NP-mPB, by adding a nucleolus-predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon-optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nucleolar organizer regions is not detected. Instead a 3-4 fold increase in piggyBac transposition efficiency is reproducibly observed in mouse and human cells. |
| format | Article |
| id | doaj-art-d8a43facd1b545b5a5f3faa0377d988e |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-d8a43facd1b545b5a5f3faa0377d988e2025-08-20T02:15:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8939610.1371/journal.pone.0089396A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells.Jin-Bon HongFu-Ju ChouAmy T KuHsiang-Hsuan FanTung-Lung LeeYung-Hsin HuangTsung-Lin YangI-Chang SuI-Shing YuShu-Wha LinChung-Liang ChienHong-Nerng HoYou-Tzung ChenPiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its potential uses in functional genomics and regenerative medicine. Efforts to increase its transposition efficiency in mammals were made through engineering the corresponding transposase (PBase) codon usage to enhance its expression level and through screening for mutant PBase variants with increased enzyme activity. To improve the safety for its potential use in regenerative medicine applications, site-specific transposition was achieved by using engineered zinc finger- and Gal4-fused PBases. An excision-prone PBase variant has also been successfully developed. Here we describe the construction of a nucleolus-predominant PBase, NP-mPB, by adding a nucleolus-predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon-optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nucleolar organizer regions is not detected. Instead a 3-4 fold increase in piggyBac transposition efficiency is reproducibly observed in mouse and human cells.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089396&type=printable |
| spellingShingle | Jin-Bon Hong Fu-Ju Chou Amy T Ku Hsiang-Hsuan Fan Tung-Lung Lee Yung-Hsin Huang Tsung-Lin Yang I-Chang Su I-Shing Yu Shu-Wha Lin Chung-Liang Chien Hong-Nerng Ho You-Tzung Chen A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. PLoS ONE |
| title | A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. |
| title_full | A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. |
| title_fullStr | A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. |
| title_full_unstemmed | A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. |
| title_short | A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells. |
| title_sort | nucleolus predominant piggybac transposase np mpb mediates elevated transposition efficiency in mammalian cells |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089396&type=printable |
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