Lentivirus vector system (lentivirus expression system, lentivirus packaging plasmid system)

Lentivirus vector system - Introduction

GeneMedi’s lentivirus Vector System, also named the lentivirus expression system or lentivirus packaging plasmid system, is a powerful tool for in-vitro & in-vivo gene delivery, shRNA mediated RNA interference (RNAi), gene editing and stable cellline development. You can easily produce a recombinant lentivirus particle in HEK293T or HEK293FT cell line in high titer using GeneMedi’s lentivirus Vector System.

The Genemedi lentivirus vector system including 3 lentivirus packaging plasmids: the lentivirus expression plasmids with different promoters and tags, an envelope protein VSV-G expressing plasmid pMD2G and a packaging plasmid pSPAX2 expressing Gag-pol and Tat.

Note:

pLV-GOI Construction

pMD2G preparation

pSPAX2 preparation


Transfection in 293T Cells


Harvesting Virus 48h and 72h post Transfection


Purification:72000g for 2 hours at 4℃


Titer Detection: Fluorescent microscopy


Transduction of Target Cells

GeneMedi has developed a variety of lentivirus expression vectors with different expression cassettes, containing kinds of verified promoters and reporters including GFP, zsgreen, RFP, mcherry and luciferase. The GeneMedi’s lentivirus expression vectors have been proved very suitalble for unique gene overexpression or shRNA-mediated knock-down (also called RNAi (RNA interference ). You can also achieve gene knock-out(KO) or gene editing using our Crispr-cas9-gRNA lentivirus expression vector.

Introduction to Lentivirus Vector System (Lentivurs packaging and expression system)

Achieve high titer of lentivirus production with GeneMedi’s lentivirus vector system (lentivirus packaging system).The lentivirus system using a low-passage HEK293T for packaging with ration of lentivirus expression vector (lentivirus overexpression or shRNA vector)and lentivirus packaging plasmid(pspax2,pMD2G)

Lentivirus are based on HIV-1 (human immunodeficiency virus type I), which has been one of the most widely used gene therapy vectors. As a powerful tool for introduction of exogenous genes, there are many advantageous features of lentivirus vectors, such as mediating efficient transfection and long-term expression of exogenous genes in both dividing and non-dividing cells. To date, lentivirus vector has been widespread utilized in various cell lines for gene overexpression, RNA interference, microRNA research and in vivo animal experiments.

A lentivirus packaging system including the lentivirus expression vector, the lentivirus package plasmid and helper plasmid. In order to meet different infection needs, Genemedi has launched a variety of lentivirus packaging system plasmids with different fluorescent labels (such as GFP, RFP, mCherry and so on), and various promoters or tags, which may also allow researchers to produce lentiviruses by themselves as they need.

Lentivirus vector system

Lentivirus expression vectors (lentivirus expression plasmids)

Note: Industry R&D excludes CRO&CDMO&CXO; Others includes CRO&CDMO&CXO&CMC&Manufacturing company

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Helper plasmids of lentivirus packaging

PMD2G: an envelope protein VSV-G expressing plasmid

Figure 1: Map and sequence of pMD2G

Click here to download SnapGene Viewer

pSPAX2: lentivirus packaging plasmids expressing Gag-pol and Tat

Figure 2: Map and sequence of pSPAX2
Click here to download SnapGene Viewer

Advantages

1. Customized cloning for any other gene ORF expression, shRNA/miRNA and CRISPR/Cas9. No known immunogenic proteins generated.

2. High titer. 108TU/ml or 109TU/ml lentiviral titer for cell line transfection in medium or large scale.

3. With wide range of host. Mediate efficient transfection in both dividing and non-dividing cells.

4. Integration into host cell genome, mediating long-term and stable expression of exogenous genes.

5. Deliver complex genetic elements, such as intron-containing sequences

6. Easy system for vector manipulation and production

Applications and Figures

Figure 1 Selected results of lentivirus infection in multiple cell lines.

Technical Documents

1. For detailed protocol about how to package and purify lentivirus, please see the pdf downloadLentivirus User Manual.

Reference

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