ArticleGene therapy2004
Engineering the splice acceptor for improved gene expression and viral titer in an MLV-based retroviral vector.
Article in Gene therapy, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- CD4(+) T Cells Modified by the Endoribonuclease MazF Are Safe and Can Persist in SHIV-infected Rhesus Macaques.Molecular therapy. Nucleic acids · 2014Article
- Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H.Gene therapy · 2014Article
- Article
- A Promising Vector for TCR Gene Therapy: Differential Effect of siRNA, 2A Peptide, and Disulfide Bond on the Introduced TCR Expression.Molecular therapy. Nucleic acids · 2012Article
- Limited complementarity between U1 snRNA and a retroviral 5' splice site permits its attenuation via RNA secondary structure.Nucleic acids research · 2009Article
- Vector design for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells.DNA repair · 2007Review
- The retrovirus RNA trafficking granule: from birth to maturity.Retrovirology · 2006Review
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have recently developed a retroviral vector that contains a splice acceptor from the human EF1-alpha gene and drives a significantly higher level of gene expression than other well known murine leukemia virus-based vectors. However, one downside of this vector is that viral titer significantly varies depending on the packaging lines used. Results from Northern blot analysis indicated that in certain cell lines the genomic transcript containing the packaging signal sequence was too efficiently spliced to the subgenomic RNA, resulting in low levels of genomic RNA and thus leading to a low viral titer. We tested the possibility of overcoming this problem by introducing mutations around the splice acceptor sequence in such a way that a delicate balance was maintained between the splicing efficiency (which determines the level of gene expression) and the amount of genomic transcript (which influences viral titer). After mutational analysis, one such mutant was found to meet this requirement. The newly constructed vector containing the engineered splice acceptor could indeed drive higher levels of expression in many therapeutic genes than other control vectors, without significantly compromising viral titer.
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Registered trials
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