Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE)

Author:

Skipper Kristian Alsbjerg,Hollensen Anne Kruse,Antoniou Michael N.,Mikkelsen Jacob GiehmORCID

Abstract

Abstract Background DNA transposon-based vectors are effective nonviral tools for gene therapy and genetic engineering of cells. However, promoter DNA methylation and a near-random integration profile, which can result in transgene integration into heterochromatin, renders such vectors vulnerable to transcriptional repression. Therefore, to secure persistent transgene expression it may be necessary to protect transposon-embedded transgenes with anti-transcriptional silencing elements. Results We compare four different protective strategies in CHO-K1 cells. Our findings show robust protection from silencing of transgene cassettes mediated by the ubiquitous chromatin-opening element (UCOE) derived from the HNRPA2B1-CBX3 locus. Using a bioinformatic approach, we define a shorter HNRPA2B1-CBX3 UCOE core fragment and demonstrate that this can robustly maintain transgene expression after extended passaging of CHO-K1 cells carrying DNA transposon vectors equipped with this protective feature. Conclusions Our findings contribute to the understanding of the mechanism of HNRPA2B1-CBX3 UCOE-based transgene protection and support the use of a correctly oriented core fragment of this UCOE for DNA transposon vector-based production of recombinant proteins in CHO-K1 cells.

Funder

Sundhed og Sygdom, Det Frie Forskningsråd

Lundbeckfonden

Hørslev-Fonden

Aase og Ejnar Danielsens Fond

Grosserer LF Foghts Fond

Agnes og Poul Friis Fond

Oda og Hans Svenningsens Fond

Snedkermester Sophus Jacobsen og Hustru Astrid Jacobsens Fond

Familien Hede Nielsens Fond

Publisher

Springer Science and Business Media LLC

Subject

Biotechnology

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