Increased in vitro and in vivo transgene expression levels mediated through cis‐acting elements

Johansen, Jens; Tornøe, Jens; Møller, Arne; Johansen, Teit E.

doi:10.1002/jgm.444

Cited by 23 publications

(17 citation statements)

References 39 publications

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“…Insulators therefore represent an attractive safety tool for retrovirus-mediated gene therapy approaches. 13,[15][16][17] In this study, we demonstrate the importance of the WPRE and insulator elements for homogeneous, stable and high level expression of CD20. Moreover, we show for the first time that CD20-transgenic T cells can efficiently be killed via RTX by activating different human effector mechanisms.…”

Section: Introductionsupporting

confidence: 53%

“…It promotes mRNA transport from the nucleus to the cytoplasm, and may facilitate protein expression from mRNAs that normally would be degraded within the nucleus. [12][13][14] In our hands, the WPRE element had only a minor positive effect on the level of CD20 expression. Surprisingly, a profound effect of the WPRE element was observed on the stability of CD20 expression in CEM cells after selection and continued culture for 3 months.…”

Section: Improved Cd20 Expression For Efficient Elimination Of Cd20-tmentioning

confidence: 91%

“…The WPRE element has been widely proven to enhance transgene expression under control of several different promoters and in different cell lineages. 13,14 Its exact mode of action is unclear, but it has been suggested that the WPRE element acts on both the transcriptional and translational level. It promotes mRNA transport from the nucleus to the cytoplasm, and may facilitate protein expression from mRNAs that normally would be degraded within the nucleus.…”

Section: Improved Cd20 Expression For Efficient Elimination Of Cd20-tmentioning

confidence: 99%

“…Improved post-transcriptional processing, transport of mRNA from the nucleus to the cytoplasm and improved translation of the mRNA have been attributed to the activity of the WPRE element. [12][13][14] Secondly, we included insulator elements in the CD20 vectors. Chicken hypersensitive site 4 (cHS4) insulators have been used in retroviral vectors for the treatment of b-chain hemoglobinopathies for stable and long-term expression of the b-globin gene.…”

Section: Introductionmentioning

confidence: 99%

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The CD20/αCD20 ‘suicide’ system: novel vectors with improved safety and expression profiles and efficient elimination of CD20-transgenic T cells

et al. 2006

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Adoptive transfer of T lymphocytes is an attractive strategy for many experimental treatment strategies for cancer. Unfortunately, manipulated T cells could be responsible for serious adverse events. Retroviral CD20-transduced T cells may be able to control these unwanted effects. CD20-positive cells are sensitive to rituximab (RTX), a monoclonal antibody specific for CD20. This permits their selective elimination in vivo in case of adverse events. To this end, a system is required that permits efficient and safe transduction of donor T cells and effective elimination of CD20-positive T cells. We constructed different CD20-encoding retroviral vectors and investigated the impact of inclusion of the woodchuck posttranscriptional regulatory element (WPRE) and the chicken hypersensitivity site 4 insulator elements on the levels, homogeneity and stability of CD20 expression. Importantly, inclusion of either WPRE or insulator elements in the retroviral vector resulted in a dramatic improvement in the stability of CD20 expression. The insulator element also led to a much more homogeneous level of CD20 expression. We also show the efficient elimination of the CD20-transgenic T cells via RTX by different effector mechanisms. In conclusion, we have constructed CD20-encoding retroviral vectors with improved efficiency and safety profiles, which can be used as a suicide strategy. Gene Therapy (2006) 13, 789-797.

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Section: Introductionsupporting

confidence: 53%

Section: Improved Cd20 Expression For Efficient Elimination Of Cd20-tmentioning

confidence: 91%

Section: Improved Cd20 Expression For Efficient Elimination Of Cd20-tmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The CD20/αCD20 ‘suicide’ system: novel vectors with improved safety and expression profiles and efficient elimination of CD20-transgenic T cells

et al. 2006

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“…To do so, it is desirable to increase the levels of transgene expression per vector genome (vg). This can be achieved through the use of genetic sequences that either increase the number of transcript copies (e.g., stronger promoters) or stabilize the mRNA (e.g., the woodchuck hepatitis virus posttranscriptional regulatory element [WPRE]) (3,16,21,22,26,29,30,32,40,48,52).…”

Section: Increased Transgene Expression Per Vector Genome Is An Impormentioning

confidence: 99%

Herpes Simplex Virus Type 1 Thymidine Kinase Sequence Fused to the lacZ Gene Increases Levels of β-Galactosidase Activity per Genome of High-Capacity but Not First-Generation Adenoviral Vectors In Vitro and In Vivo

Puntel

Barrett

Mondkar

et al. 2009

J Virol

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Prospects and implications of using chromatin insulators in gene therapy and transgenesis

2004

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Gene therapy has emerged from the idea of inserting a wild‐type copy of a gene in order to restore the proper expression and function of a damaged gene. Initial efforts have focused on finding the proper vector and delivery method to introduce a corrected gene to the affected tissue or cell type. Even though these first attempts are clearly promising, seveal problems remain unsolved. A major problem is the influence of chromatin structure on transgene expression. To overcome chromatin‐dependent repressive transgenic states, researchers have begun to use chromatin regulatory elements to drive transgene expression. Insulators or chromatin boundaries are able to protect a transgene against chromatin position effects at their genomic integration sites, and they are able to maintain transgene expression for long periods of time. Therefore, these elements may be very useful tools in gene therapy applications for ensuring high‐level and stable expression of transgenes. BioEssays 26:796–807, 2004. © 2004 Wiley Periodicals, Inc.

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Increased in vitro and in vivo transgene expression levels mediated through cis‐acting elements

Abstract: The data suggest that incorporating cis-acting elements in gene therapy vectors may result in improvements to currently available therapeutic vectors.

Cited by 23 publications

References 39 publications

The CD20/αCD20 ‘suicide’ system: novel vectors with improved safety and expression profiles and efficient elimination of CD20-transgenic T cells

The CD20/αCD20 ‘suicide’ system: novel vectors with improved safety and expression profiles and efficient elimination of CD20-transgenic T cells

Herpes Simplex Virus Type 1 Thymidine Kinase Sequence Fused to the lacZ Gene Increases Levels of β-Galactosidase Activity per Genome of High-Capacity but Not First-Generation Adenoviral Vectors In Vitro and In Vivo

Prospects and implications of using chromatin insulators in gene therapy and transgenesis

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