Description
pLVX-DsRed-Monomer-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to DsRed-Monomer, a monomeric mutant of the Discosoma sp. red fluorescent protein. Genes cloned into the multiple cloning site (MCS),located upstream of the DsRed-Monomer coding sequence, are expressed as N-terminal fusions of the DsRed-Monomer protein. Expression of the fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of DsRed-Monomer fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses DsRed-Monomer, and may be used to produce marker virus to
optimize infection protocols.
pLVX-DsRed-Monomer-N1 contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and nhances nuclear export of viral and transgene RNA (1), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-DsRed-Monomer-N1 also contains a central polypurine tract (cPPT) element that increases nuclear importation of the viral genome during target cell infection, resulting in improved vector integration and more efficient transduction (3).
In addition to lentiviral elements, pLVX-DsRed-Monomer-N1 contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
Use
pLVX-DsRed-Monomer-N1 constitutively expresses your gene of interest from PCMV IE when transduced into target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging cells with our Lenti-X™ HTX Packaging System (Cat. Nos. 631247 and 631249). This packaging system allows you to safely produce high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4).
