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Authors: Scheuermann, Barbara
Diem, Tanja
Ivics, Zoltán
Andrade, Miguel
Title: Evolution-guided evaluation of the inverted terminal repeats of the synthetic transposon Sleeping Beauty
Online publication date: 23-Apr-2019
Year of first publication: 2019
Language: english
Abstract: 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.
DDC: 570 Biowissenschaften
570 Life sciences
Institution: Johannes Gutenberg-Universität Mainz
Department: FB 10 Biologie
Place: Mainz
Version: Published version
Publication type: Zeitschriftenaufsatz
License: CC BY
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Journal: Scientific reports
Pages or article number: Art. 1171
Publisher: Macmillan Publishers Limited, part of Springer Nature
Publisher place: London
Issue date: 2019
ISSN: 2045-2322
Publisher URL:
Publisher DOI: 10.1038/s41598-018-38061-w
Annotation: Andrade, Miguel veröffentlicht unter: Andrade-Navarro, Miguel A.
Appears in collections:JGU-Publikationen

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