Retroviral vectors, by nature, infect and stably integrate into the genome of the host cell. Therefore, any cell line expressing the fluorescent protein marker from a retroviral vector is already an established stable cell line, unlike those from plasmid constructs. Coexpression of genes from different promoters can lead to competition between the promoters, and the eventual repression of one of the promoter elements. To eliminate this problem, our vectors use a single promoter to drive the production of IRES-containing bicistronic transcripts. The IRES allows cap-independent translation from an internal start site, resulting in stable coexpression of your protein of interest along with a fluorescent marker protein (Mountford and Smith, 1995). For expressing fluorescently tagged fusion proteins, use Retro-X Living Colors fusion vectors with DsRed-Monomer or AcGFP1.
- Gene editing
- Viral transduction
- T-cell transduction and culture
- Inducible systems
- Transfection reagents
- Fluorescent proteins
Fluorescent retroviral expression vectors
- Efficient delivery of fusion proteins into hard-to-transfect cells
- Enhanced gene expression
- High-titer, self-inactivating retroviral backbone ensures robust expression
Our Retro-X IRES Living Colors vectors are designed to efficiently coexpress your gene of interest with our brightest red (DsRed-Express) or green (ZsGreen1) fluorescent markers. This is accomplished through the production of bicistronic transcripts that contain an internal ribosome entry site (IRES). The IRES allows independent translation of a second protein from a single transcript (Figure 1). These vectors contain all of the features necessary to efficiently deliver your gene of interest into even hard-to-transfect cells.
Additional benefits to using these vectors include detection of the fluorescent marker by flow cytometry or fluorescence microscopy (Figure 2), allowing you to easily determine your transfection/transduction efficiency. Second, because both proteins are translated from a single transcript, the expression level of the easily detectable fluorescent marker allows you to approximate that of your gene of interest. Third, the fluorescent marker allows you to use flow cytometry to enrich for cells expressing your gene of interest. This is especially useful when introducing your gene of interest into hard-to-transfect cells (Figure 3.) Finally, you can rapidly titer the infectivity of your viral stocks by counting fluorescent cells or by using flow cytometry.
Our vectors are based on the pMIN series of retroviral vectors, noted for their high titers, enhanced gene expression, and improved safety profiles (Yu et al. 2000). High titer production and enhanced gene expression result from the presence of highly optimized splicing machinery, and the MMLV LTR promoter (Figure 4). Improved safety profiles, on the other hand, result from the removal of all retroviral coding sequences, which reduces the likelihood that replication competent retrovirus will result from homologous recombination (Yu et al. 2000).
Retro-X Living Colors Fusion vectors are designed to facilitate delivery of distinct red and green fluorescent proteins that have been specifically engineered to provide outstanding performance when expressed as a fusion with your protein of interest (Gurskaya et al. 2003).
DsRed-Monomer and AcGFP1 proteins are ideal tools for monitoring gene expression and intracellular protein trafficking. Because of their distinct spectra, these fluorescent proteins can be used for multicolor labeling and direct visualization applications. Furthermore, due to their monomeric character, it is now possible to localize AcGFP1 and DsRed Monomer to compartments and structures that can only be targeted with non-oligomerizing fusion tags. Both DsRed-Monomer and AcGFP1 proteins are extremely stable and are ideal for subcellular localization studies. With these vectors, you can visualize biological processes as they occur and also easily track your protein of interest to a specific subcellular organelle or structure.
These vectors offer advantages beyond those of standard plasmids. Retroviral vectors, by nature, infect and stably integrate into the genome of the host cell. Therefore, any cell line expressing the fluorescent protein marker is already an established stable cell line. Constructing dual-color stable cell lines has never been easier (Figure 5).
Retro-X IRES Living Colors vectors are designed to efficiently coexpress your gene of interest with DsRed-Express or ZsGreen1 fluorescent markers. Retro-X Living Colors fusion vectors contain all of the features necessary to efficiently deliver your fluorescently tagged protein of interest into hard-to-transfect cell types. Both N- and C-terminal versions of the DsRed Monomer and AcGFP1 fluorescent proteins are available.
These vectors allow for easy detection of the fluorescent marker by flow cytometry or fluorescence microscopy to determine your transfection/transduction efficiency. Since both proteins are translated from a single transcript, the expression level of the easily detectable fluorescent marker allows you to approximate that of your gene of interest. In addition, the fluorescent marker allows you to use flow cytometry to enrich for cells expressing your gene of interest. This is especially useful when introducing your gene of interest into hard-to-transfect cells. Finally, you can rapidly titer the infectivity of your viral stocks by counting fluorescent cells or by using flow cytometry.
Clontech-brand fluorescent protein vectors: now only $149
Clontech (now Takara Bio USA, Inc.) was the first to bring GFP to market. We commercialized mCherry, DsRed, and tdTomato. We've generated vectors in every color of the rainbow, and now we're bringing you a spectrum of savings. From now through March 31st, you can get any of over 170 fluorescent protein vectors for only $149—just pick your application and choose your vectors.
Cite Discount Quote 35732 when ordering to receive the discount. Offer valid in the US and Canada only through 03.31.20. No other discounts apply.Offer details Vector information
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