Evaluation of Tat-Encoding Bicistronic Human Immunodeficiency Virus Type 1 Gene Transfer Vectors in Primary Canine Bone Marrow Mononuclear Cells

Srinivasakumar, Narasimhachar; Zaboikin, Michail; Zaboikina, Tatiana; Schuening, Friedrich

doi:10.1128/jvi.76.14.7334-7342.2002

Cited by 3 publications

(2 citation statements)

References 37 publications

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“…The remnant portion of the HIV-1 env contains the RRE. The vector has an engineered frame-shift (FS) mutation in gag [ 6 ] and the central polypurine tract and central termination sequences (CPPT/CTS) to improve gene-transfer efficiency [ 23 - 25 ]. The transgene expression cassette, positioned between the BamHI site in the second coding exon of Rev that overlaps the 3' end of env and the XhoI site in nef , consists of human elongation factor 1 alpha (EF1α) promoter driving enhanced green fluorescent protein (EGFP).…”

Section: Methodsmentioning

confidence: 99%

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Srinivasakumar

2008

AIDS Res Ther

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Background: Human immunodeficiency virus type 1 (HIV-1)-based gene delivery systems are popular due to their superior efficiency of transduction of primary cells. However, these systems cannot be readily used for delivery of anti-HIV-1 genes that target constituents of the packaging system itself due to inimical effects on vector titer. Here we describe HIV-1-based packaging systems containing the Rev-response element (RRE), of simian immunodeficiency virus (SIV) in place of the HIV-1 RRE. The SIV RRE-containing packaging systems were used to deliver the antiRev gene, Rev M10, into HIV-1 susceptible target cells.

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Section: Methodsmentioning

confidence: 99%

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Srinivasakumar

2008

AIDS Res Ther

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“…To create pN-GIT72-1xCTE, the CPPT/CTS sequence from pN-GIT72/CPPT [38] was isolated using BssHII and BsaBI and used to replace the corresponding sequence in pN-GITC [27] . To create pN-GIT72-2xCTE, a NotI-XhoI fragment, containing EMCV-IRES and one copy of CTE, was isolated from pN-GIT72-1xCTE and ligated into pN-GIT72-1xCTE digested with NotI and SalI which cuts at the 5′ end of the CTE thus resulting in a vector containing two copies of CTE.…”

Section: Methodsmentioning

confidence: 99%

Rev-Free HIV-1 Gene Delivery System for Targeting Rev-RRE-Crm1 Nucleocytoplasmic RNA Transport Pathway

Srinivasakumar

2011

PLoS ONE

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The use of RNA transport elements from different viruses can provide novel attributes to HIV-1-based gene delivery systems such as improved safety or Rev independence. We previously described an HIV-1 based gene delivery system that utilized the simian immunodeficiency virus Rev-response element (RRE) in place of the HIV-1 RRE. Despite the use of Rev for the production of vector stocks, we showed the utility of this system for delivery of Rev M10, a dominant-negative mutant of HIV-1 Rev, into T-cells. Here, we investigated the use of RNA transport elements from Mason-Pfizer monkey virus or MPMV for the creation of high-titered Rev-free HIV-1-based packaging systems. The HIV-1 gag/pol expression constructs containing one or more copies of MPMV constitutive RNA transport element (CTE) were used to package similarly modified gene-transfer vectors in the presence or absence of Rev. An inverse correlation between the number of CTE modules and Rev dependency was noted for vector stock production. While packaging systems containing multiple CTEs were resistant to exogenously expressed Rev M10, the titers of vectors encoding Rev M10 were nevertheless reduced in comparison to vectors encoding only green fluorescent protein (GFP). In contrast, a gene transfer vector encoding the Rev M10 transgene and containing both RNA transport elements exhibited almost no loss in titer in comparison to a corresponding vector encoding only GFP. The optimized Rev-independent gene delivery system was used for delivery of Rev M10 transgene into T-lymphocytes. Upon challenge in single round infection assays with HIV-1, the modified T-cells produced fewer virus particles than control cells expressing GFP. This Rev-free packaging system may prove useful for targeting the Rev-RRE-Crm1 nucleocytoplasmic RNA transport pathway for inhibiting HIV replication.

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Cloning and Expression of CanineO⁶-Methylguanine-DNA Methyltransferase in Target Cells, Using Gammaretroviral and Lentiviral Vectors

et al. 2004

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The human O(6)-methylguanine-DNA methyltransferase (MGMT) gene and its mutants have been used for in vivo selection of transduced hematopoietic stem cells with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) alone or in combination with O(6)-benzylguanine (BG). To allow similar in vivo selection in dogs, without the risk of inducing an immune response, we have cloned the canine MGMT drug resistance gene. Comparison of canine and human MGMT-coding regions indicates that there is about 62% amino acid identity and 78% similarity between the two MGMTs. The canine MGMT is also longer, by nine amino acids. Proline at position 140 and the surrounding amino acids of the human MGMT are highly conserved in the canine sequence. To determine whether mutation of the proline residue at position 144 to lysine in the canine MGMT would provide a similar advantage for selection of transduced cells as the human mutant, Moloney murine leukemia virus and human immunodeficiency type 1 vectors encoding the corresponding mutant MGMT were created and used to express separately canine and human MGMTs in cultured cells. Drug resistance assays using BCNU alone or BCNU with BG demonstrated that the wild-type and mutant canine MGMTs provided resistance to the selection agents that was comparable to the human MGMT counterparts.

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Evaluation of Tat-Encoding Bicistronic Human Immunodeficiency Virus Type 1 Gene Transfer Vectors in Primary Canine Bone Marrow Mononuclear Cells

Cited by 3 publications

References 37 publications

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Rev-Free HIV-1 Gene Delivery System for Targeting Rev-RRE-Crm1 Nucleocytoplasmic RNA Transport Pathway

Cloning and Expression of CanineO⁶-Methylguanine-DNA Methyltransferase in Target Cells, Using Gammaretroviral and Lentiviral Vectors

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Evaluation of Tat-Encoding Bicistronic Human Immunodeficiency Virus Type 1 Gene Transfer Vectors in Primary Canine Bone Marrow Mononuclear Cells

Cited by 3 publications

References 37 publications

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Substitution of the Rev-response element in an HIV-1-based gene delivery system with that of SIVmac239 allows efficient delivery of Rev M10 into T-lymphocytes

Rev-Free HIV-1 Gene Delivery System for Targeting Rev-RRE-Crm1 Nucleocytoplasmic RNA Transport Pathway

Cloning and Expression of CanineO6-Methylguanine-DNA Methyltransferase in Target Cells, Using Gammaretroviral and Lentiviral Vectors

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Cloning and Expression of CanineO⁶-Methylguanine-DNA Methyltransferase in Target Cells, Using Gammaretroviral and Lentiviral Vectors