Biosynthetic retrovectoring systems for gene therapy

Hodgson, Clague P.; Xu, Guoping; Solaiman, Fauzia; Zink, Mary Ann

doi:10.1007/s001090050110

Cited by 11 publications

(6 citation statements)

References 32 publications

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“…By eliminating the redundancy of restriction enzyme recognition sites found in dual LTR vectors, it was possible to insert alternative promoter sequences into LTR promoter cassettes (Hodgson et al, 1997). However, transcription from single LTR vectors was limited by their unusual structure, in which one LTR in a circular vector acts as both the 5Ј-and 3Ј-LTRs.…”

Section: Single Ltr Vectors For Manipulation Of Thementioning

confidence: 99%

“…Reverse transcriptase PCR (RT-PCR) was used to isolate the U3 (promoter) sequences from the VL30 RNA from several tissues expressing them, and seamless cloning was used to recombine these U3 sequences with the LTRs in single LTR vectors (Hodgson et al, 1997(Hodgson et al, , 1998b. A liver-derived LTR was introduced into Hep G2 cells, expressing a green fluorescent protein reporter gene.…”

Section: Incorporating Tissue-specific Ltrs Intomentioning

confidence: 99%

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Modular retro-vectors for transgenic and therapeutic use

Solaiman

Zink

et al. 2000

Mol. Reprod. Dev.

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Section: Single Ltr Vectors For Manipulation Of Thementioning

confidence: 99%

Section: Incorporating Tissue-specific Ltrs Intomentioning

confidence: 99%

Modular retro-vectors for transgenic and therapeutic use

Solaiman

Zink

et al. 2000

Mol. Reprod. Dev.

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“…There are three components to improve efficiency, the first of which is to improve gene transfer efficiency. Many efforts have been made to increase the infectious titer of recombinant viral vectors (Hodgson et al, 1997), and new nonviral molecules have been developed for more efficient delivery of foreign genes (Ledley, 1995). We have developed novel viral liposomes by com bining fusion proteins derived from Sendai virus (HVJ; hemagglutinating virus of Japan) with DNA-containing liposom es (Kaneda et al, 1989a), and we created more efficient viral liposomes by changing the lipid components of the liposomes (Saeki et al, 1997).…”

Section: Introduction T O a D V A N Ce H U M A N G En E Th E Ra Pymentioning

confidence: 99%

Enhancement of Transgene Expression by Cotransfection of oriP Plasmid with EBNA-1 Expression Vector

Kaneda

Saeki²,

Nakabayashi³

et al. 2000

Human Gene Therapy

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We have attempted to develop a system for specific enhancement of transgene expression, which has been one of the most important issues in human gene therapy. When an Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1) expression vector, pCMV-trEBNA-1, was cotransfected with an origin of latent viral DNA replication (oriP)-harboring plasmid, poriP-CMV-luciferase, luciferase gene expression was up to 20 times greater than in the absence of EBNA-1. This enhancement was regulated mainly at the transcriptional level and was dependent on the oriP sequence and the amount of EBNA-1. However, cointroduction of poriP-CMV-luciferase with purified recombinant EBNA-1 inhibited luciferase gene expression whereas no inhibition was observed when pCMV-luciferase was cointroduced with recombinant EBNA-1. We also introduced poriP-CMV-luciferase into mouse liver via the use of HVJ (hemagglutinating virus of Japan)-liposomes. By 10 days after transfer, luciferase gene expression was decreased to low levels. We then introduced pCMV-trEBNA-1 to mouse liver via HVJ-liposomes on day 10. Luciferase gene expression was reactivated, whereas no reactivation was detected by the injection of EBNA-1 expression plasmid into liver injected with pCMV-luciferase lacking the oriP sequence. Thus, cotransfection of oriP-harboring expression vector with EBNA-1 expression plasmid should be promising for human gene therapy, although the safety of the system must be investigated thoroughly.

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“…The commonly used DNA transfection procedures are (a) chemical methods, including calcium phosphate precipitation, DEAE-dextran complexation and lipid-mediated DNA transfer; (b) physical methods, including electroporation, microinjection, and biolistic particle delivery; and (c) recombinant viral vectors (1,2). The majority of previous investigations have focused on elucidating the function of the transfected genes.…”

Section: Introductionmentioning

confidence: 99%

A method to monitor DNA transfer during transfection

et al. 1999

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This report describes a method for monitoring the transfer of DNA during transfection. This method involves random labeling of plasmid DNA with fluorescein-12-dUTP, flow cytometric detection and sorting of the fluorescent transfected cells, and laser confocal fluorescence microscopic determination of the intracellular location of the plasmid DNA. By this method, >95% of the sorted cells were labeled, indicating a >95% specificity of the sorting procedure. The sorted cells were viable, as indicated by their ability to exclude trypan blue dye (>98% cells excluded the dye) and to maintain cell growth. The results of the kinetics of the Lipofectin transfection technology show that the fraction of the cells that internalized plasmid DNA increased from 10% at 1 hr after initiation of the transfection procedures to 18% at 3 hr. This method does not require protein expression, does not require the use of selection pressure such as drug treatment to isolate the cells that internalized DNA, and can be used to study the early events of DNA transfection.

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Biosynthetic retrovectoring systems for gene therapy

Cited by 11 publications

References 32 publications

Modular retro-vectors for transgenic and therapeutic use

Modular retro-vectors for transgenic and therapeutic use

Enhancement of Transgene Expression by Cotransfection of oriP Plasmid with EBNA-1 Expression Vector

A method to monitor DNA transfer during transfection

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