cDNA Expression Cell Lines and Constructs

Lentiviral expression vectors are one of the most effective vehicles to introduce and stably express cDNA, as well as other genetic elements (e.g., sgRNA, shRNA, etc.) into almost any mammalian cell, including non-dividing cells and whole model organisms. Lentiviral expression constructs packaged into pseudoviral particles can be transduced into cells with very high efficiency, approaching 100% in some cell types. Packaged lentiviral constructs can be transduced into even the most difficult to transfect cells, such as primary, stem, and differentiated cells, with high efficiency.

Cellecta offers both lentiviral cDNA cloning vectors, including our InDOXible Tet-Activated cDNA Vector System, and custom-made cDNA expression constructs, as well as related products--such as the PromoterTest Kit--to help ensure you get optimal expression of your gene of interest—and reagents for packaging and transduction.

    • Large range of lentiviral constructs with various selection genes (e.g., PuroR, BleoR, NeoR, Hyg) fluorescent markers (GFP, RFP), and promoters (e.g., CMV, EFS, UbiC, RSV).
    • Our InDOXible Tet-Activated cDNA System provides options for inducible cDNA expression.
    • All lentiviral constructs are fully sequenced for quality control and provide as plasmid or pre-packaged viral particles.
    • We will design and make custom cDNA expression constructs for you, or engineer protein expression cell lines.
  • A range of constitutive and tet-activated inducible cDNA cloning vectors are available off-the-shelf for you to insert your gene of interest. These can be ordered directly through our website, or by other standard means. If you can’t find a vector design you need, please contact us. Since we are use these systems for in-house projects, we frequently have variations not shown on the website.

    For our lentiviral cDNA construction service, you simply need to provide

    • the sequence of the cDNA you would like expressed
    • your choice of one of our vectors, or specify the selection markers and promoters desired

    Cellecta will then

    • Synthesize the cDNA
    • Clone the cDNA into the desired vector
    • Sequence it to ensure proper construction

    Cellecta can provide you the cDNA construct--in either plasmid or packaged formats--to introduce into your cells, or we can make the expression cell line for you.

  • Lentiviral expression vectors offer and convenient and efficient approach to introduce a cDNA sequence stably into the genomic DNA of virtually any mammalian cells. Lentiviral vectors have the genetic elements required for packaging, transduction, stable integration of the viral expression construct into genomic DNA, and expression of a gene of interest. Since transduced lentiviral vectors integrate into the genomic DNA, the cDNA they carry is heritable as the cells propagate.

    High titer VSV-G pseudoviral particles are produced by transiently co-transfecting into producer cells (e.g., HEK293 cells) the expression construct and packaging plasmids, which provide the proteins essential for transcription and packaging of an RNA copy of lentiviral vector into recombinant pseudoviral particles. High titer vesicular stomatitis virus (VSV-G) pseudotyped viral particles are produced and secreted by the co-transfected packaging cells into the culture media where they can be harvested to transduce (i.e., infect) target cells with the expression constructs.

    The VSV-G pseudotyped viral particles efficiently mediate viral entry through lipid binding and plasma membrane fusion to infect virtually any mammalian cells, as well as some avian cell lines. The lentiviral vectors are engineered so that they do not have the required elements to make viral particles without the associated proteins on the packaging plasmids. As a result, once they are introduced into the target cells, they do not generate virus.

    Pseudotyped lentiviruses have been successfully used to transduce many other cell types, including neuronal, dendritic, endothelial, retinal, pancreatic, hepatic, aortic smooth muscle cells, airway epithelia, skin fibroblasts, macrophages, etc. Lentivectors have been successfully used also for direct in vivo delivery and expression of transgenes in muscle, brain, airway epithelium, liver, pancreas, retina, and skin.

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