dc.identifier.uri |
http://dx.doi.org/10.15488/16160 |
|
dc.identifier.uri |
https://www.repo.uni-hannover.de/handle/123456789/16287 |
|
dc.contributor.author |
Tschorn, Natalie
|
|
dc.contributor.author |
van Heuvel, Yasemin
|
|
dc.contributor.author |
Stitz, Jörn
|
|
dc.date.accessioned |
2024-02-08T08:59:00Z |
|
dc.date.available |
2024-02-08T08:59:00Z |
|
dc.date.issued |
2022 |
|
dc.identifier.citation |
Tschorn, N.; van Heuvel, Y.; Stitz, J.: Transgene Expression and Transposition Efficiency of Two-Component Sleeping Beauty Transposon Vector Systems Utilizing Plasmid or mRNA Encoding the Transposase. In: Molecular Biotechnology 65 (2022), S. 1327-1335. DOI: https://doi.org/10.1007/s12033-022-00642-6 |
|
dc.description.abstract |
The use of two-component transposon plasmid vector systems, namely, a transposase construct and a donor vector carrying the gene of interest (GOI) can accelerate the development of recombinant cell lines. However, the undesired stable transfection of the transposase construct and the sustained expression of the enzyme can cause genetic instability due to the re-mobilization of the previously transposed donor vectors. Using a Sleeping Beauty-derived vector system, we established three recombinant cell pools and demonstrate stable integration of the transposase construct and sustained expression of the transposase over a period of 48 days. To provide an alternative approach, transcripts of the transposase gene were generated in vitro and co-transfected with donor vector plasmid at different ratios and mediating high GOI copy number integrations and expression levels. We anticipate that the use of transposase mRNA will foster further improvements in future cell line development processes. |
eng |
dc.language.iso |
eng |
|
dc.publisher |
New York, NY : Springer |
|
dc.relation.ispartofseries |
Molecular Biotechnology 65 (2022) |
|
dc.rights |
CC BY 4.0 Unported |
|
dc.rights.uri |
https://creativecommons.org/licenses/by/4.0 |
|
dc.subject |
mRNA transfection |
eng |
dc.subject |
Producer cell line |
eng |
dc.subject |
Sleeping Beauty |
eng |
dc.subject |
Transgene expression |
eng |
dc.subject |
Transposition |
eng |
dc.subject |
Transposon vector |
eng |
dc.subject.ddc |
570 | Biowissenschaften, Biologie
|
|
dc.subject.ddc |
540 | Chemie
|
|
dc.subject.ddc |
660 | Technische Chemie
|
|
dc.title |
Transgene Expression and Transposition Efficiency of Two-Component Sleeping Beauty Transposon Vector Systems Utilizing Plasmid or mRNA Encoding the Transposase |
eng |
dc.type |
Article |
|
dc.type |
Text |
|
dc.relation.essn |
1559-0305 |
|
dc.relation.issn |
1073-6085 |
|
dc.relation.doi |
https://doi.org/10.1007/s12033-022-00642-6 |
|
dc.bibliographicCitation.volume |
65 |
|
dc.bibliographicCitation.firstPage |
1327 |
|
dc.bibliographicCitation.lastPage |
1335 |
|
dc.description.version |
publishedVersion |
eng |
tib.accessRights |
frei zug�nglich |
|