Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty
Sleeping Beauty (SB) is a synthetic Tc1/mariner transposon that is widely used for genetic engineering in vertebrates, including humans. Its sequence was derived from a consensus of sequences found in fish species including the Atlantic salmon (Salmo salar). One of the functional components of SB, t...
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ftpubmed:oai:pubmedcentral.nih.gov:6362248 2023-05-15T15:32:22+02:00 Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty Scheuermann, Barbara Diem, Tanja Ivics, Zoltán Andrade-Navarro, Miguel A. 2019-02-04 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362248/ http://www.ncbi.nlm.nih.gov/pubmed/30718656 https://doi.org/10.1038/s41598-018-38061-w en eng Nature Publishing Group UK http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362248/ http://www.ncbi.nlm.nih.gov/pubmed/30718656 http://dx.doi.org/10.1038/s41598-018-38061-w © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. CC-BY Article Text 2019 ftpubmed https://doi.org/10.1038/s41598-018-38061-w 2019-02-10T01:43:46Z Sleeping Beauty (SB) is a synthetic Tc1/mariner transposon that is widely used for genetic engineering in vertebrates, including humans. Its sequence was derived from a consensus of sequences found in fish species including the Atlantic salmon (Salmo salar). One of the functional components of SB, the transposase enzyme, has been subject to extensive mutagenesis yielding hyperactive protein variants for advanced applications. The second functional component, the transposon inverted terminal repeats (ITRs), has so far not been extensively modified, mainly due to a lack of natural sequence information. Importantly, as genome sequences become available, they can provide a rich source of information for a refined molecular definition of the functional components of these transposons. Here we have mined the Salmo salar genome for a comprehensive set of transposon sequences that were used to build a refined consensus sequence. We synthetically produced the new consensus ITR sequences and used them to build a new transposon, the performance of which has been tested in cell-based transposition assays. The consensus sequence did not support enhanced transposition, suggesting alternative mechanisms responsible for the preferential amplification of these sequence variants in the salmon genome. Text Atlantic salmon Salmo salar PubMed Central (PMC) Scientific Reports 9 1 |
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Article Scheuermann, Barbara Diem, Tanja Ivics, Zoltán Andrade-Navarro, Miguel A. Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
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Article |
description |
Sleeping Beauty (SB) is a synthetic Tc1/mariner transposon that is widely used for genetic engineering in vertebrates, including humans. Its sequence was derived from a consensus of sequences found in fish species including the Atlantic salmon (Salmo salar). One of the functional components of SB, the transposase enzyme, has been subject to extensive mutagenesis yielding hyperactive protein variants for advanced applications. The second functional component, the transposon inverted terminal repeats (ITRs), has so far not been extensively modified, mainly due to a lack of natural sequence information. Importantly, as genome sequences become available, they can provide a rich source of information for a refined molecular definition of the functional components of these transposons. Here we have mined the Salmo salar genome for a comprehensive set of transposon sequences that were used to build a refined consensus sequence. We synthetically produced the new consensus ITR sequences and used them to build a new transposon, the performance of which has been tested in cell-based transposition assays. The consensus sequence did not support enhanced transposition, suggesting alternative mechanisms responsible for the preferential amplification of these sequence variants in the salmon genome. |
format |
Text |
author |
Scheuermann, Barbara Diem, Tanja Ivics, Zoltán Andrade-Navarro, Miguel A. |
author_facet |
Scheuermann, Barbara Diem, Tanja Ivics, Zoltán Andrade-Navarro, Miguel A. |
author_sort |
Scheuermann, Barbara |
title |
Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
title_short |
Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
title_full |
Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
title_fullStr |
Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
title_full_unstemmed |
Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty |
title_sort |
evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon sleeping beauty |
publisher |
Nature Publishing Group UK |
publishDate |
2019 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362248/ http://www.ncbi.nlm.nih.gov/pubmed/30718656 https://doi.org/10.1038/s41598-018-38061-w |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362248/ http://www.ncbi.nlm.nih.gov/pubmed/30718656 http://dx.doi.org/10.1038/s41598-018-38061-w |
op_rights |
© The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
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CC-BY |
op_doi |
https://doi.org/10.1038/s41598-018-38061-w |
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