Lentivirus Vectors Encoding Both Central Polypurine Tract and Posttranscriptional Regulatory Element Provide Enhanced Transduction and Transgene Expression

Barry, Simon C.; Harder, Brandon; Brzezinski, Margaret; Flint, Lisa; Seppen, Jurgen; Osborne, William

doi:10.1089/104303401750214311

Cited by 130 publications

(119 citation statements)

References 28 publications

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“…The vector used in this study is a third-generation, self-inactivating (SIN) vector, with major part of the 3 0 U3 region deleted, including the TATA box. 27 The Rev-responsive element (RRE), 28 the central polypurine tract (cPPT) [29][30][31] and the human hepatitis B virus-derived posttranscriptional regulatory element (PRE) are indicated. The encephalomyocardin virus internal ribosomal entry site (IRES) was obtained from pTM3.…”

Section: Efficient Incorporation Of Pix_tcr A1m1 In the Virus Capsidmentioning

confidence: 99%

Adenovirus targeting to HLA-A1/MAGE-A1-positive tumor cells by fusing a single-chain T-cell receptor with minor capsid protein IX

et al. 2008

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Section: Efficient Incorporation Of Pix_tcr A1m1 In the Virus Capsidmentioning

confidence: 99%

Adenovirus targeting to HLA-A1/MAGE-A1-positive tumor cells by fusing a single-chain T-cell receptor with minor capsid protein IX

et al. 2008

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“…48 For some transfer vectors, the human CMV promoter was replaced with the murine stem cell virus promoter (MSCV) 49 or the CK6 promoter. 25 VSV-G-pseudotyped lentiviral vectors were produced by cotransfecting 293T cells, and purified by ultracentrifugation as described.…”

Section: Construction Of Lentiviral Transfer Vectors and Viral Preparmentioning

confidence: 99%

“…25 VSV-G-pseudotyped lentiviral vectors were produced by cotransfecting 293T cells, and purified by ultracentrifugation as described. 48 The titer of viruses containing reporter genes was measured by transducing Hela cells with serial dilutions of vector preparations. The titer of other vector stocks was estimated by measuring viral p24 gag antigen using the HIV-1 p24 Antigen Assay kit (Beckman Coulter Inc.) and comparing to the titer of vectors with reporter genes as described.…”

Section: Construction Of Lentiviral Transfer Vectors and Viral Preparmentioning

confidence: 99%

“…The titer of other vector stocks was estimated by measuring viral p24 gag antigen using the HIV-1 p24 Antigen Assay kit (Beckman Coulter Inc.) and comparing to the titer of vectors with reporter genes as described. 48 Single myofiber isolation and culture TA muscles were digested with 0.2% Collagenase II (Worthington Biochemical Corporation) for 40 min. Single myofibers were obtained by pipetting up and down using a Pasture pipette and then cultured in MethoCult GF M3534 medium (StemCell Technologies).…”

Section: Construction Of Lentiviral Transfer Vectors and Viral Preparmentioning

confidence: 99%

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Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin

Kimura

Fall

et al. 2005

Gene Ther

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Gene therapy for Duchenne muscular dystrophy (DMD) will require sustained expression of therapeutic dystrophins in striated muscles. Lentiviral vectors have a relatively large transgene carrying capacity and can integrate into nondividing cells. We therefore explored the use of lentiviral vectors for transferring genes into mouse skeletal muscle cells. These vectors successfully transferred a minidystrophin expression cassette into mdx muscles, and minidystrophin expression persisted and prevented subsequent muscle fiber degeneration for at least 6 months. However, only low to moderate levels of skeletal muscle transduction could be obtained by intramuscular injection of the highest currently available lentiviral doses. Using cultured cells, the lentiviral vectors effectively transduced proliferating and terminally differentiated muscle cells, indicating that cell cycling is not essential for transduction of myogenic cells. We further showed that lentiviral vectors efficiently transduced both primary myoblasts and multipotent adult progenitor cells (MAPCs) in vitro, and the cells persistently expressed transgenes without any obvious toxicity. When mdx primary myoblasts were genetically modified with minidystrophin vectors and transplanted into mdx skeletal muscles, significant numbers of dystrophin-expressing myofibers formed. Finally, we showed that a short, highly active CK6 regulatory cassette directed muscle-specific activity in the context of the lentiviral vectors. The ability of lentiviral vectors to transduce myogenic progenitors using a minidystrophin cassette regulated by a muscle-specific promoter suggests that this system could be useful for ex vivo gene therapy of muscular dystrophy.

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“…19,21 Plasmids pRRL-cPPT-CMV-GFP-PRE-SIN (here named pLV-CMV-GFP), pRRL-cPPT-CMV-LUC-PRE-SIN (here named pLV-CMV-Luc), and pRRL-cPPT-CMV-IRES-GFP (here named pLV-CMV-IRES-GFP) were constructed with cytomegalovirus promoter driving green fluorescent protein or luciferase reporter genes directly or indirectly via an Internal Ribosome Entry Site (IRES). Then, plasmids pRRL-cPPT-CMV-CC8 (here named pLV-CMV-CC8), pRRL-cPPT-CMV-FKC8 (here named pLV-CMV-FKC8), and pRRL-cPPT-CMV-CC8-IRES-GFP (here named pLV-CMV-CC8-IRES-GFP) and pRRL-cPPT-CMV-FKC8-IRES-GFP (here named pLV-CMV-FKC8-IRES-GFP) were generated by insertion of CC8 or FKC8 inserts in pLV-CMV or pLV-CMV-IRES-GFP constructs, respectively.…”

Section: Lentiviruses Construction Production and Transductionmentioning

confidence: 99%

Development of an inducible suicide gene system based on human caspase 8

et al. 2005

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Suicide gene-therapy strategies are promising approaches in treating various diseases such as cancers, atherosclerosis, and graft-versushost-disease. Here, we describe the development of a new effector gene based on inducing functional caspase 8, the initiator caspase in the death-receptor pathway. We constructed vectors encoding a constitutively active form of human caspase 8 (CC8), and demonstrated the efficient killing of a variety of cell types in transfection and lentivirus-transduction assays. We then analyzed the ability to control the apoptotic activity of a caspase 8-derived construct through the ARIADt homodimerization system (FKC8), a system shown to be extremely effective in several cellular models upon retroviral and lentiviral gene transfer. Similarly, two transcription-regulation systems, muristerone-regulated and Tet-On, were tested to control the expression of CC8. The homodimerization-regulated system FKC8 was shown to be the most efficient system with low background activity in noninduced conditions. In the presence of a dimerizer, it was as active as the activated Tet-On system. From our data, we conclude that the dimerizer-dependent human caspase 8 represents a highly inducible and very powerful system to eradicate transduced cell populations. In addition to its application in experimental gene therapy, this variant may be highly useful for mechanistic research related to apoptosis.

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Lentivirus Vectors Encoding Both Central Polypurine Tract and Posttranscriptional Regulatory Element Provide Enhanced Transduction and Transgene Expression

Cited by 130 publications

References 28 publications

Adenovirus targeting to HLA-A1/MAGE-A1-positive tumor cells by fusing a single-chain T-cell receptor with minor capsid protein IX

Adenovirus targeting to HLA-A1/MAGE-A1-positive tumor cells by fusing a single-chain T-cell receptor with minor capsid protein IX

Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin

Development of an inducible suicide gene system based on human caspase 8

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