Get lentiviral constructs or engineered cell lines expressing your cDNA of interest.
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 is able to make cDNA expression constructs for your use, or engineer and provide you with protein expression cell lines.
For our lentiviral cDNA construct 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 cell line, or we can make the expression cell line for you.
Lentiviral expression vectors contain the genetic elements required for packaging, transduction, stable integration of the viral expression construct into genomic DNA, and expression of the shRNA, sgRNA, cDNA, or reporter. 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. Lentiviral packaging plasmids provide all of the proteins essential for transcription and packaging of an RNA copy of the expression construct into recombinant pseudoviral particles. Following transfection, the expression construct transcript is efficiently packaged into vesicular stomatitis virus (VSV-G) pseudotyped viral particles. The pseudoviral particles secreted by producer cells in culture media are used to transduce expression constructs into target cells.
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. Pseudotyped lentiviruses have been successfully used to infect 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.