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dc.contributor.authorSchambach, Axel
dc.contributor.authorBohne, J
dc.contributor.authorChandra, Saurabh
dc.contributor.authorWill, Elke
dc.contributor.authorMargison, Geoffrey P
dc.contributor.authorWilliams, David A
dc.contributor.authorBaum, Christopher
dc.date.accessioned2009-07-09T12:24:18Z
dc.date.available2009-07-09T12:24:18Z
dc.date.issued2006-02
dc.identifier.citationEqual potency of gammaretroviral and lentiviral SIN vectors for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells. 2006, 13 (2):391-400 Mol. Ther.en
dc.identifier.issn1525-0016
dc.identifier.pmid16226060
dc.identifier.doi10.1016/j.ymthe.2005.08.012
dc.identifier.urihttp://hdl.handle.net/10541/73118
dc.description.abstractSevere adverse events related to insertional mutagenesis have reinforced interest in self-inactivating (SIN) retroviral vectors lacking enhancer-promoter sequences in the long terminal repeats (LTRs). Here, we have compared the potency of gammaretroviral and lentiviral vectors expressing the P140K mutant of O(6)-methylguanine-DNA methyltransferase (MGMT). MGMT-P140K is a clinically relevant selection marker that mediates a strong survival advantage in hematopoietic cells exposed to alkylating agents. We designed gammaretroviral and lentiviral vectors that contained identical enhancer-promoter sequences located either in the LTR or downstream of the packaging region, for internal initiation of transcription from SIN backbones. Gammaretroviral vectors with intact LTRs containing enhancer-promoter sequences showed both higher titers and higher expression levels than the lentiviral counterparts, likely a result of suboptimal RNA processing of the lentiviral leader region. In the SIN context, gammaretroviral and lentiviral vectors with comparable internal cassettes had similar expression properties. Interestingly, gammaretroviral SIN vectors pseudotyped with RD114/TR had a higher transduction efficiency on proliferating human CD34(+) cells than lentiviral counterparts. These results encourage further investigations into the formation of retroviral hybrid vectors that combine the desired properties of high efficiency and increased biosafety.
dc.language.isoenen
dc.subjectHaematopoietic Stem Cellsen
dc.subjectLeukaemiaen
dc.subject.meshAnimals
dc.subject.meshCells, Cultured
dc.subject.meshGene Expression Regulation, Viral
dc.subject.meshGenetic Vectors
dc.subject.meshHematopoietic Stem Cells
dc.subject.meshHumans
dc.subject.meshLentivirus
dc.subject.meshLeukemia Virus, Murine
dc.subject.meshMice
dc.subject.meshMice, Inbred C57BL
dc.subject.meshMutagenesis, Insertional
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase
dc.subject.meshRNA Processing, Post-Transcriptional
dc.subject.meshTransduction, Genetic
dc.titleEqual potency of gammaretroviral and lentiviral SIN vectors for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells.en
dc.typeArticleen
dc.contributor.departmentDepartment of Hematology, Hemostaseology, and Oncology, Hannover Medical School, Germany.en
dc.identifier.journalMolecular Therapyen
html.description.abstractSevere adverse events related to insertional mutagenesis have reinforced interest in self-inactivating (SIN) retroviral vectors lacking enhancer-promoter sequences in the long terminal repeats (LTRs). Here, we have compared the potency of gammaretroviral and lentiviral vectors expressing the P140K mutant of O(6)-methylguanine-DNA methyltransferase (MGMT). MGMT-P140K is a clinically relevant selection marker that mediates a strong survival advantage in hematopoietic cells exposed to alkylating agents. We designed gammaretroviral and lentiviral vectors that contained identical enhancer-promoter sequences located either in the LTR or downstream of the packaging region, for internal initiation of transcription from SIN backbones. Gammaretroviral vectors with intact LTRs containing enhancer-promoter sequences showed both higher titers and higher expression levels than the lentiviral counterparts, likely a result of suboptimal RNA processing of the lentiviral leader region. In the SIN context, gammaretroviral and lentiviral vectors with comparable internal cassettes had similar expression properties. Interestingly, gammaretroviral SIN vectors pseudotyped with RD114/TR had a higher transduction efficiency on proliferating human CD34(+) cells than lentiviral counterparts. These results encourage further investigations into the formation of retroviral hybrid vectors that combine the desired properties of high efficiency and increased biosafety.


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