Next Article in Journal
Essential Oils Biofilm Modulation Activity, Chemical and Machine Learning Analysis—Application on Staphylococcus aureus Isolates from Cystic Fibrosis Patients
Previous Article in Journal
Adiponectin Role in Neurodegenerative Diseases: Focus on Nutrition Review
 
 
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon

by
Kevin He
,
S. M. Ali Hosseini Rad
*,
Aarati Poudel
and
Alexander Donald McLellan
*
Department of Microbiology and Immunology, University of Otago, Dunedin 9016, New Zealand
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(23), 9256; https://doi.org/10.3390/ijms21239256
Submission received: 27 August 2020 / Revised: 26 November 2020 / Accepted: 2 December 2020 / Published: 4 December 2020
(This article belongs to the Section Molecular Biology)

Abstract

Promoter choice is an essential consideration for transgene expression in gene therapy. The expression of multiple genes requires ribosomal entry or skip sites, or the use of multiple promoters. Promoter systems comprised of two separate, divergent promoters may significantly increase the size of genetic cassettes intended for use in gene therapy. However, an alternative approach is to use a single, compact, bidirectional promoter. We identified strong and stable bidirectional activity of the RPBSA synthetic promoter comprised of a fragment of the human Rpl13a promoter, together with additional intron/exon structures. The Rpl13a-based promoter drove long-term bidirectional activity of fluorescent proteins. Similar results were obtained for the EF1-α and LMP2/TAP1 promoters. However, in a lentiviral vector, the divergent bidirectional systems failed to produce sufficient titres to translate into an expression system for dual chimeric antigen receptor (CAR) expression. Although bidirectional promoters show excellent applicability to drive short RNA in Sleeping Beauty transposon systems, their possible use in the lentiviral applications requiring longer and more complex RNA, such as dual-CAR cassettes, is limited.
Keywords: Sleeping Beauty transposon; bidirectional promoters; gene expression; gene therapy; synthetic biology; RPBSA; EF-1α; LMP2/TAP1 Sleeping Beauty transposon; bidirectional promoters; gene expression; gene therapy; synthetic biology; RPBSA; EF-1α; LMP2/TAP1

Share and Cite

MDPI and ACS Style

He, K.; Rad, S.M.A.H.; Poudel, A.; McLellan, A.D. Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon. Int. J. Mol. Sci. 2020, 21, 9256. https://doi.org/10.3390/ijms21239256

AMA Style

He K, Rad SMAH, Poudel A, McLellan AD. Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon. International Journal of Molecular Sciences. 2020; 21(23):9256. https://doi.org/10.3390/ijms21239256

Chicago/Turabian Style

He, Kevin, S. M. Ali Hosseini Rad, Aarati Poudel, and Alexander Donald McLellan. 2020. "Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon" International Journal of Molecular Sciences 21, no. 23: 9256. https://doi.org/10.3390/ijms21239256

APA Style

He, K., Rad, S. M. A. H., Poudel, A., & McLellan, A. D. (2020). Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon. International Journal of Molecular Sciences, 21(23), 9256. https://doi.org/10.3390/ijms21239256

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop