Abrogated cryptic activation of lentiviral transfer vectors

Luche, Ralf M.; Enssle, Joerg; Kiem, Hans Peter

doi:10.1038/srep00438

Cited by 2 publications

(2 citation statements)

References 29 publications

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“…37 We are developing integration-deficient LVs for use as vaccines in immunotherapy. We were able to demonstrate that design changes resulting in the RI gag/pol vector did not affect its in vivo potency compared with WT gag/pol vector for both integration-competent and integration-deficient versions.…”

Section: Discussionmentioning

confidence: 99%

“…Since the origin of third-generation LVs, additional attempts have been made at enhancing the predictable safety profiles of vectors for example either by separating gag/pol components 36 or by conditional removal of the packaging signal from proviral DNA. 37 We are developing integration-deficient LVs for use as vaccines in immunotherapy. We were able to demonstrate that design changes resulting in the RI gag/pol vector did not affect its in vivo potency compared with WT gag/pol vector for both integration-competent and integration-deficient versions.…”

Section: Discussionmentioning

confidence: 99%

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A Rev-Independentgag/polEliminates Detectable psi-gag Recombination in Lentiviral Vectors

Tareen

Nicolai

Campbell

et al. 2013

BioResearch Open Access

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Lentiviral vectors (LVs) are being developed for clinical use in humans for applications including gene therapy and immunotherapy. A safety concern for use of LVs in humans is the generation of replication-competent lentivirus (RCL), which may arise due to recombination between the split genomes of third-generation LVs. Although no RCL has been detected to date, design optimizations that minimize recombination events between split genome vectors would provide an added safety benefit that may further reduce the risk of RCL formation. Here we describe design elements introduced to the gag/pol plasmid with the intention of eliminating psi-gag recombination between the vector genome and gag/pol. These design changes, consisting of codon optimization of the gag/pol sequence and the deletion of the Rev-responsive element, abrogate the requirement for Rev in expression of Gag protein, thus the resulting gag/pol construct being Rev independent (RI gag/pol). We show that generating vector using the RI gag/pol construct has no effect on particle production or transduction titers. The RI and wild-type gag/pol vectors function equivalently as antigen-specific immunotherapy, potently inducing antigen-specific CD8 T cells that protect against challenge with vaccinia virus. Most importantly, the designed RI gag/pol eliminated detectable psi-gag recombination. Interestingly, we detected recombination between the vector genome and gag/pol from regions without sequence homology. Our findings imply that although unpredictable recombination events may still occur, the RI gag/pol design is sufficient to prevent psi-gag recombination.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A Rev-Independentgag/polEliminates Detectable psi-gag Recombination in Lentiviral Vectors

Tareen

Nicolai

Campbell

et al. 2013

BioResearch Open Access

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Over-Expression of Deubiquitinating Enzyme USP14 in Lung Adenocarcinoma Promotes Proliferation through the Accumulation of β-Catenin

Bai

et al. 2013

IJMS

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The deubiquitinating enzyme USP14 has been identified and biochemically studied, but its role in lung cancer remains to be elucidated. The aim of this study was to evaluate the prognostic significance of USP14 in patients with lung adenocarcinoma and to define its role in lung cancer cell proliferation. USP14 mRNA levels in different non-small cell lung cancer (NSCLC) cell lines were detected by real-time qPCR. USP14 protein levels in surgically resected samples from NSCLC patients, and in NSCLC cell lines, were detected by immunohistochemistry or Western blot. The correlation of USP14 expression with clinical characteristics and prognosis was determined by survival analysis. After silencing USP14, cell proliferation was assessed by MTT assay and the cell cycle was measured by FACS assay. It was found that USP14 expression was upregulated in NSCLC cells, especially in adenocarcinoma cells. Over-expression of USP14 was associated with shorter overall survival of patients. Downregulation of USP14 expression arrested the cell cycle, which may be related to β-catenin degradation. Over-expression of USP14 was associated with poor prognosis in NSCLC patients and promoted tumor cell proliferation, which suggests that USP14 is a tumor-promoting factor and a promising therapeutic target for NSCLC.

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Abrogated cryptic activation of lentiviral transfer vectors

Cited by 2 publications

References 29 publications

A Rev-Independentgag/polEliminates Detectable psi-gag Recombination in Lentiviral Vectors

A Rev-Independentgag/polEliminates Detectable psi-gag Recombination in Lentiviral Vectors

Over-Expression of Deubiquitinating Enzyme USP14 in Lung Adenocarcinoma Promotes Proliferation through the Accumulation of β-Catenin

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