TY  - JOUR
U1  - Wissenschaftlicher Artikel
A1  - Tschorn, Natalie
A1  - van Heuvel, Yasemin
A1  - Stitz, Jörn
T1  - Transgene Expression and Transposition Efficiency of Two-Component Sleeping Beauty Transposon Vector Systems Utilizing Plasmid or mRNA Encoding the Transposase
JF  - Molecular Biotechnology
N2  - 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.
KW  - Transposition
KW  - Transposon Vector
KW  - mRNA Transfection
KW  - Transgene Expression
KW  - Producer Cell Line
Y1  - 2022
UN  - https://nbn-resolving.org/urn:nbn:de:hbz:832-epub4-27620
SN  - 1073-6085
SS  - 1073-6085
SN  - 1559-0305
SS  - 1559-0305
U6  - https://doi.org/10.1007/s12033-022-00642-6
DO  - https://doi.org/10.1007/s12033-022-00642-6
VL  - 65
IS  - 8
SP  - 1327
EP  - 1335
S1  - 9
PB  - Springer US
ER  -