Use of a murine secreted alkaline phosphatase as a non‐immunogenic reporter gene in mice

Mælandsmo, Gunhild Mari; Ross, P. Joel; Pavliv, Marta; Meulenbroek, Robert A.; Evelegh, Carole; Muruve, Daniel A.; Graham, Frank L.; Parks, Robin J.

doi:10.1002/jgm.666

Cited by 26 publications

(32 citation statements)

References 32 publications

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“…This is in marked contrast to the rapid decrease in SEAP activity observed in huSEAP-treated animals. The difference in expression of human and mouse SEAP indicates that huSEAP clearance occurs through an immune-mediated mechanism, which is consistent with previous observations showing that the plasma huSEAP level is in inverse proportion to the anti-huSEAP antibody titer (15,29,42). As noted, the strongest SEAP expression was attained with conventional promoters, and not with the MCK/SV40 (L) promoter, which might be a reflection of the net promoter activity for a nonimmunogenic reporter gene.…”

Section: Discussionsupporting

confidence: 89%

Muscle creatine kinase/SV40 hybrid promoter for muscle-targeted long-term transgene expression

Takeshita

Takase²,

Tozuka³

et al. 2007

Int J Mol Med

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Abstract. Gene therapy for congenital protein deficiencies requires lifelong expression of a deficient protein. Current gene therapy approaches preferentially employ the strong cytomegalovirus (CMV) promoter/enhancer or its derivative CAG promoter; however, these promoters provide only temporary transgene expression. To create a promoter that enables long-lasting expression in muscle, hybrid promoters were constructed by coupling the muscle creatine kinase (MCK) enhancer to various strong promoters for enhancement of tissue specificity and improved transcriptional activity. A hybrid promoter containing the MCK enhancer and the simian virus 40 promoter (MCK/SV40 promoter) yielded long-term (>6 months) expression of a human secretory alkaline phosphatase (huSEAP) reporter gene following electrotransfer of the plasmid into mice, whereas expression using a conventional CMV or CAG promoter faded away within a few weeks. To explore the mechanism behind the sustained expression obtained with the MCK/SV40 promoter, mice were immunized with a LacZ expression plasmid driven by MCK/SV40 or a conventional promoter. Minimal cellular and humoral responses to LacZ were observed in MCK/SV40 promotertreated animals, and mouse SEAP gene expression in vivo was successfully maintained by both the MCK/SV40 and conventional promoters. These results suggest that the lower immunogenicity of the MCK/SV40 promoter contributed to long-lasting gene expression in mice. Therefore, the MCK/ SV40 promoter may provide the basis for development of an effective transgene expression cassette for treatment of congenital protein deficiencies in which therapeutic proteins are recognized as foreign by the host immune system.

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Section: Discussionsupporting

confidence: 89%

Muscle creatine kinase/SV40 hybrid promoter for muscle-targeted long-term transgene expression

Takeshita

Takase²,

Tozuka³

et al. 2007

Int J Mol Med

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“…Thus, pRL6 and the resulting virus, AdRL6, encode the pIX-MR1 fusion protein replacing the native pIX coding sequence within the Ad genome and under regulation by the native pIX promoter. A BamHI/MfeI fragment from pRP2202, containing the murine secreted alkaline phosphatase (mSEAP) reporter gene under regulation by the mouse phosphoglycerate kinase promoter and simian virus 40 (SV40) polyadenylation sequence (43), was used to replace a BamHI/MfeI fragment in pRL5, generating pRL7, which was subsequently recovered as an infectious plasmid designated pRL34. AdRP2218, an E1/E3-deleted Ad that contains an mSEAP expression cassette identical to that of pRL34 and an unmodified pIX gene, has been described previously (43,49).…”

Section: Methodsmentioning

confidence: 99%

Retargeting of Adenovirus Vectors through Genetic Fusion of a Single-Chain or Single-Domain Antibody to Capsid Protein IX

Poulin

Lanthier

Smith

et al. 2010

J Virol

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Adenovirus (Ad) vectors are the most commonly used system for gene therapy applications, due in part to their ability to infect a wide array of cell types and tissues. However, many therapies would benefit from the ability to target the Ad vector only to specific cells, such as tumor cells for cancer gene therapy. In this study, we investigated the utility of capsid protein IX (pIX) as a platform for the presentation of single-chain variable-fragment antibodies (scFv) and single-domain antibodies (sdAb) for virus retargeting. We show that scFv can be displayed on the capsid through genetic fusion to native pIX but that these molecules fail to retarget the virus, due to improper folding of the scFv. Redirecting expression of the fusion protein to the endoplasmic reticulum (ER) results in correct folding of the scFv and allows it to recognize its epitope; however, ER-targeted pIX-scFv was incorporated into the Ad capsid at a very low level which was not sufficient to retarget virus infection. In contrast, a pIX-sdAb construct was efficiently incorporated into the Ad capsid and enhanced virus infection of cells expressing the targeted receptor. Taken together, our data indicate that pIX is an effective platform for presentation of large targeting polypeptides on the surface of the virus capsid, but the nature of the ligand can significantly affect its association with virions.

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“…14) To determine whether SEAP expression can be monitored when delivered with a nonviral vector, we injected plasmid DNAs containing the Seap gene, driven by the CMV and EF1α promoters. We used a hydrodynamics-based administration method that enables the efficient introduction of naked DNA into mouse liver.…”

Section: Resultsmentioning

confidence: 99%

“…Maelandsmo et al designed an adenoviral vector containing the gene encoding secreted alkaline phosphatase (SEAP), derived from the murine FVB strain. 14) They injected the virus into FVB mice, and found that the SEAP protein was non-immunogenic. Meanwhile, Wolff et al inserted the target sequence of miR142-3p, a microRNA expressed in cells of hematopoietic origin, into the region corresponding to the 3′ untranslated region (UTR) of the luciferase gene.…”

mentioning

confidence: 99%

Effects of Endogenous Proteins and microRNA Target Sequence in a Positive Feedback System

Kanda

Togashi

Harashima

et al. 2012

Biological & Pharmaceutical Bulletin

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A positive feedback system, using GAL4-vp16 (a fusion protein of yeast GAL4 and herpes simplex virus vp16) as an activator and firefly luciferase as a reporter, maintained luciferase expression for 7 d in mice. However, the luciferase expression decreased after 7 d, and this phenomenon could be caused by immunoreactions against these exogenous proteins. This hypothesis was examined by the following three strategies, designed to avoid the putative immunoreactions: (i) use of the endogenous secreted alkaline phosphatase (SEAP) protein as a reporter, (ii) replacement of vp16 with endogenous transcription factors, and (iii) insertion of the target sequence of microRNA expressed in cells of hematopoietic origin, to suppress GAL4-vp16 expression in antigen-presenting cells. The results obtained in this study suggested that silencing would be induced by mechanism(s) besides immunoreactions against reporter and activator proteins.

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Use of a murine secreted alkaline phosphatase as a non‐immunogenic reporter gene in mice

Abstract: Taken together, these data illustrate that mSEAP is a sensitive, non-immunogenic reporter gene for preclinical mouse studies.

Cited by 26 publications

References 32 publications

Muscle creatine kinase/SV40 hybrid promoter for muscle-targeted long-term transgene expression

Muscle creatine kinase/SV40 hybrid promoter for muscle-targeted long-term transgene expression

Retargeting of Adenovirus Vectors through Genetic Fusion of a Single-Chain or Single-Domain Antibody to Capsid Protein IX

Effects of Endogenous Proteins and microRNA Target Sequence in a Positive Feedback System

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