A novel Flk1-TVA transgenic mouse model for gene delivery to angiogenic vasculature

Vervoort, Virginie; Lu, Mark; Valencia, Fatima; Lesperance, Jacqueline; Breier, Georg; Oshima, Robert G.; Pasquale, Elena B.

doi:10.1007/s11248-007-9156-1

Cited by 6 publications

(3 citation statements)

References 38 publications

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“…This results in TVA expression in the developing embryonic vasculature, subcutaneous implants in adult mice, and tumor-associated endothelial cells in adult mice. Limited infection of tumor-associated vasculature was showed when this TVA line was combined with the MMTV-PyMT mammary tumor model (Vervoort et al 2008). This mouse model may be a useful tool in investigating gene perturbations that influence the function of tumor-associated vasculature.…”

Section: Tumor Models Using Rcas-tvamentioning

confidence: 99%

“…One such example is active endothelium, which forms blood vessels to support tumor formation. A mouse model expressing TVA under the Flk1 promoter results in TVA expression in proliferating endothelial cells (Vervoort et al 2008). This results in TVA expression in the developing embryonic vasculature, subcutaneous implants in adult mice, and tumor-associated endothelial cells in adult mice.…”

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Using the RCAS-TVA System to Model Human Cancer in Mice

Ahronian

Lewis

2014

Cold Spring Harb Protoc

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For successful infection, avian sarcoma leukosis virus subgroup A (ASLV-A) requires its receptor, tumor virus A (TVA), to be present on the surface of target cells. This is the basis of the RCAS-TVA gene delivery system: Mammalian cells lack the gene encoding TVA and are normally resistant to infection by ASLV; however, transgenic targeting of TVA to specific cell types or tissues in the mouse renders these cells uniquely susceptible to infection by ASLV-A-based RCAS viruses. The RCAS-TVA system is a powerful tool for effectively modeling human tumors, including pancreatic, ovarian, and breast cancers, gliomas, and melanomas. RCAS viruses can deliver cDNAs (≤2.8 kb), as well as short hairpin RNAs (shRNAs), microRNAs (miRNAs), and other noncoding RNAs. Compared with traditional transgenic and knockout mice, the RCAS-TVA system has several strengths. First, virus delivery is generally performed postnatally and results in a relatively low infection rate of target cells; the sporadic postnatal expression of the gene of interest mimics the situation in developing human tumors. Second, a single transgenic mouse line can be used to compare the consequences of specific genes on tumor development, with viruses encoding oncogenes or shRNAs targeting specific tumor suppressor genes. TVA mouse strains can also be easily combined with transgenic, knock-in, and knockout mouse models to study cooperating genetic events. THE RCAS VIRUSRCAS stands for replication-competent avian sarcoma-leukosis virus (ASLV) long-terminal repeat (LTR) with splice acceptor. The virus enters cells via the TVA receptor, which mammalian cells do not natively express. Following entry into a mammalian cell, a cDNA copy of the viral genome is made. When the cell undergoes division, the viral cDNA copy incorporates into the mammalian host genome.The RCAS virus retains all of the necessary viral genes for replication. Therefore, in an avian cell culture, there is no need for the use of helper plasmids to generate virus as is commonly done with retroviruses that have tropism for mammalian cells. The immortalized chicken fibroblast cell line DF1 is commonly used to propagate RCAS viruses in culture (Himly et al. 1998;Schaefer-Klein et al. 1998). Chicken embryo fibroblasts may also be used. In addition, high-titer virus can be produced for several days or weeks following a single transfection of a viral plasmid into avian producer cells. (See Protocol: Generation of High-Titer RCAS Virus from DF1 Chicken Fibroblasts [Ahronian and Lewis 2014].) The virus glycoprotein binds to its receptor to gain entry into a cell. Because only avian cells naturally express the TVA receptor to the RCAS glycoprotein, in the absence of engineered TVA expression infection of mammalian cells is very limited. Inserts in the RCAS retrovirus are stable up to 2.8 kb but less stable when >2.8 kb. It is common beyond this insert size that RCAS will lose its insert, generate truncation mutants, or produce low-titer virus. The limited carrying capacity of RCAS viruses is one of the majo...

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Section: Tumor Models Using Rcas-tvamentioning

confidence: 99%

mentioning

confidence: 99%

Using the RCAS-TVA System to Model Human Cancer in Mice

Ahronian

Lewis

2014

Cold Spring Harb Protoc

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“…There have been several studies reporting the application of this TVA-RCAS system in non-avian organisms, mostly in mice [5,21]. In 1994, Federspiel et al were the first to generate a transgenic mouse, in which the TVA receptor was under the control of the promoter of the chicken gene skeletal muscle α-actin [5].…”

Section: Introductionmentioning

confidence: 99%

Selective cell targeting and lineage tracing of human induced pluripotent stem cells using recombinant avian retroviruses

Hildebrand

Seemann

Kurtz

et al. 2015

Cell. Mol. Life Sci.

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Human induced pluripotent stem cells (hiPSC) differentiate into multiple cell types. Selective cell targeting is often needed for analyzing gene function by overexpressing proteins in a distinct population of hiPSC-derived cell types and for monitoring cell fate in response to stimuli. However, to date, this has not been possible, as commonly used viruses enter the hiPSC via ubiquitously expressed receptors. Here, we report for the first time the application of a heterologous avian receptor, the tumor virus receptor A (TVA), to selectively transduce TVA(+) cells in a mixed cell population. Expression of the TVA surface receptor via genetic engineering renders cells susceptible for infection by avian leucosis virus (ALV). We generated hiPSC lines with this stably integrated, ectopic TVA receptor gene that expressed the receptor while retaining pluripotency. The undifferentiated hiPSC(TVA+) as well as their differentiating progeny could be infected by recombinant ALV (so-called RCAS virus) with high efficiency. Due to incomplete receptor blocking, even sequential infection of differentiating or undifferentiated TVA(+) cells was possible. In conclusion, the TVA/RCAS system provides an efficient and gentle gene transfer system for hiPSC and extends our possibilities for selective cell targeting and lineage tracing studies.

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The RCAS/TVA Somatic Gene Transfer Method in Modeling Human Cancer

Ferris

Lewis

et al. 2011

Genetically Engineered Mice for Cancer Research

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A novel Flk1-TVA transgenic mouse model for gene delivery to angiogenic vasculature

Cited by 6 publications

References 38 publications

Using the RCAS-TVA System to Model Human Cancer in Mice

Using the RCAS-TVA System to Model Human Cancer in Mice

Selective cell targeting and lineage tracing of human induced pluripotent stem cells using recombinant avian retroviruses

The RCAS/TVA Somatic Gene Transfer Method in Modeling Human Cancer

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