pPACKH1 HIV Lentivector Packaging KitUser Manual
- Generate high titers of transduction-ready pseudoviral particles
- Efficiently deliver and stably express your lentivector constructs in a wide range of mammalian cells
- Quantify pseudoviral titers and transduction efficiency using a convenient copGFP control
- 293TN producer cell line optimized for high pseudoviral titers also available
Designed to package any of SBI’s RNAi, PathNet, or cDNA Cloning and Expression Lentivectors into pantropic pseudoviral particles, the HIV-based pPACK Packaging System makes it easy to deliver and express lentivector constructs in a wide-range of mammalian cells.
Generate transducible lentiviral particles with your expression construct.
The pPACK Packaging Plasmid Mix, available separately for FIV and HIV-based systems, produces all the structural and replication proteins needed to generate VSV-G-pseudotyped lentiviral particles containing an expression construct (constructed using an SBI Lentiviral CRISPR or RNAi vector, PathNet™ reporter, or cDNA vector). The pPACK Packaging Plasmid Mix enables the cells to replicate expression constructs and package them into pseudoviral particles.
When pPACK Plasmids are co-transfected into HEK 293T cells (see link below) with an expression construct that contains the genetic elements required for packaging, transduction, and expression, the cells (called “packaging" or “producer" cells) efficiently package the expression construct into highly transducible VSV-G-pseudotyped lentiviral particles.
Highly efficient delivery of any lentivector construct.
You can use pseudoviral particles produced by packaging cells to introduce the expression construct into any mammalian cells—including dividing and quiescent cells, primary cells, stem cells, neuronal cells, endothelial cells, etc. Pseudoviral transduction is much more efficient and widely applicable than standard transfection. Once delivered, the lentivector expression construct integrates into the genomic DNA and heritably expresses the siRNA sequence.
Achieve high titers with the 293TN producer cell line.
To facilitate packaging, we offer SBI's 293TN producer cell line which is optimized for effective production of pseudoviral particles at high titers, see link below. The 293TN cells stably express the SV40 large T antigen and neomycin gene products.
Co-transfect HEK293T cells with pHIV-PACK plasmid mix and your pHIV expression product and produce transduction-ready pseudoviral particles
200 ul pPACKH1 Packaging Plasmid Mix: Mixture of pPACKH1-GAG, pPACKH1-
REV, and pVSV-G plasmids (0.5 ug/µl)
20 ul pSIH1-H1-copGFP Positive Control Expression Plasmid (0.5 ug/µl)
Please note: The HIV-based Packaging Kit is also available in the Lentisuite Deluxe Kit including HEK293TN Cell Line, PEG-It Lentivirus Precipitation Solution and Ultrarapid Lentivirus Titer Kit.