Moloney murine sarcoma proviral DNA is a transcriptional unit

Benz, Edmund W.; Wydro, Robert; Nadal‐Ginard, Bernardo; Dina, Dino

doi:10.1038/288665a0

Cited by 94 publications

(47 citation statements)

References 45 publications

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“…The viral TATA box and following sequences were not replaced in these vectors, since the terminal bases of U3 are necessary for recognition of the polyadenylation signal in the 3Ј LTR. 24 Virus harvested from bulk populations of stable TE-FLY-A8 transfectants was used to infect PAE and TE671 target cells. Endogenous expression in PAE cells was confirmed by RT-PCR using primers specific for porcine ppET1, whereas no endogenous expression was detected in TE671 cells (data not shown) that were subsequently used as a negative control.…”

Section: Generation Of Retroviral Vectors With Hybrid Ltr By Replacemmentioning

confidence: 99%

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

2000

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Tumour growth is dependent upon a blood supply and is associated with the switch to the angiogenic phenotype. We are developing strategies for targeting gene expression to endothelial cells in the tumour vasculature. Recombinant retroviruses have been generated that incorporate regulatory sequences of the prepro-endothelin-1 (ppET1) promoter. Following reverse transcription and integration these modifications are duplicated in the proviral 5Ј LTR for transcription of the internal ␤-galactosidase reporter gene. The titres and endothelial specificity of retroviral vectors harbouring different modifications have been analysed. In the optimal strategy, replacing the MLV enhancer with ppET1 promoter sequences containing the GATA and AP1 elements whilst

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Section: Generation Of Retroviral Vectors With Hybrid Ltr By Replacemmentioning

confidence: 99%

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

2000

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“…lA), leads to the functional difference between the retroviral 5' and 3' poly(A) sites (Benz et al 1980;Dougherty and Temin 1987). However, we have shown that this hypothesis is unlikely (Iwasaki and Temin 1990).…”

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confidence: 97%

The efficiency of RNA 3'-end formation is determined by the distance between the cap site and the poly(A) site in spleen necrosis virus.

Iwasaki

Temin²

1990

Genes & Development

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The efficiency of RNA 3'-end formation of spleen necrosis virus (SNV) is determined by the distance between the cap site and the poly(A) site. When the distance between the cap site and the poly(A) site was shorter than 500 bases, only 3-9% of the RNA was polyadenylated at the SNV poly(A) site. However, when the distance between the cap site and the poly(A) site was 1400 bases or more, 70% of the total RNA was polyadenylated at the SNV poly(A] site. In contrast, the poly(A) signal sequences of the thymidine kinase {tk) and SV40 late genes functioned at high efficiency, even with a distance between the cap site and the poly(A) site that was short enough to inactivate the SNV poly(A) signal. Therefore, this distance-dependent inactivation of RNA 3'-end formation is specific for SNV sequences and perhaps for related retroviruses. This finding explains the difference between the 5' and 3' poly(A) sites in many retrovirus RNAs.

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“…3,1983 on May 12, 2018 by guest http://mcb.asm.org/ Downloaded from was cleaved with PstI, which excised the major central region (approximately 3.95 kb) of the Mo-MSV genome. This left intact the 5' and 3' LTR sequences and short lengths of Mo-MSV sequences immediately adjacent to the LTRs, whose significance will be discussed later.…”

Section: Resultsmentioning

confidence: 99%

“…There is evidence that the origin of certain rapidly transforming murine retroviruses, for example, Moloney murine sarcoma virus (Mo-MSV), occurs by the recombination of Moloney murine leukemia virus (MoMuLV) with a cellular gene or oncogene (c-mos) normally present in the mouse genome (17,27). The gene now carried by the virus (designated vmos) becomes integrated, apparently randomly, into the genome of cells infected with Mo-MSV, where it is expressed at a high level and thus leads to cell transformation (3,12). Except for Rous sarcoma virus, the rapidly transforming retroviruses are replication defective, but they can be packaged in virion coats in the presence of a suitable helper retrovirus (43).…”

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confidence: 99%

Design of a Retrovirus-Derived Vector for Expression and Transduction of Exogenous Genes in Mammalian Cells

et al. 1983

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We have developed a transfection vector for animal cells that contains long terminal repeat (LTR) sequences to promote expression. Plasmid p101/101, a derivative of plasmid pBR322 containing the complete Moloney murine sarcoma virus genome, was cut with restriction enzymes and religated so that both the 5′ and 3′ LTRs were retained and all but about 700 base pairs of the intervening viral sequences were removed. To test this vector, theEscherichia coligenegptwas cloned into a uniquePstI site, between the two LTRs, with guanine and cytosine tailing, a method that can be generalized for insertion of any DNA segment into this vector. When DNA from recombinant plasmids in which thegptgene was inserted in the same transcriptional polarity as the LTR sequences was transfected onto murine or rat fibroblast cultures, we obtained a high yield of Gpt+colonies. However, plasmid constructs with thegptgene in the opposite polarity were virtually devoid of activity. Withgptin the proper orientation, restriction enzyme cuts within the LTRs or between the 5′ LTR and thegptgene reduced transfection by more than 98%, whereas a cut between thegptgene and the 3′ LTR gave an 80% reduction in activity. Thus, both 5′ and 3′ LTR sequences are essential for optimalgptexpression, although the 5′ LTR appears to play a more important role. When the LTR-gptplasmid was transfected onto murine leukemia virus-infected mouse fibroblasts, we obtained evidence that RNA copies became pseudotyped into viral particles which could transfer the Gpt+phenotype into rodent cells with extremely high efficiency. This vector should prove useful for high-efficiency transduction of a variety of genes in mammalian cells.

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Moloney murine sarcoma proviral DNA is a transcriptional unit

Cited by 94 publications

References 45 publications

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

Generation of a high titre retroviral vector for endothelial cell-specific gene expression in vivo

The efficiency of RNA 3'-end formation is determined by the distance between the cap site and the poly(A) site in spleen necrosis virus.

Design of a Retrovirus-Derived Vector for Expression and Transduction of Exogenous Genes in Mammalian Cells

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