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Lentiviral Technology

Expression Vectors:

Safe, stable and effective gene delivery
The human (HIV) and feline (FIV) Lentiviral Vector Systems provide exceptional tools to express any sequence in virtually all mammalian cells. The system consists of three main components:
(1) the lentiviral expression plasmids (also known as "transfer vector")
(2) the lentiviral packaging plasmid mix
(3) a producer cell line for packaging

To create functional pseudoviral particles, the lentivector containing the expression construct is co-transfected with the packaging plasmids into the producer cell line. The packaging cells replicate the expression construct and package it into pseudoviral particles. These pseudoviral particles are then purified and used to deliver the expression construct to the cells of interest. Once delivered, the viral expression construct integrates, and the construct is expressed. SBI’s lentivectors are self-inactivating. Upon integration into the genome, the 5’ LTR promoter is inactivated, which prevents formation of replication-competent viral particles. Thus, the transduced cell cannot produce additional viral particles since the cell does not contain the genes needed to produce the viral capsid, nor is there a functional 5’LTR to enable it to replicate the viral sequence.

In addition to yielding very high expression levels, the lentiviral system is very effective at delivering genetic material to almost any mammalian cell—including non-dividing cells and whole model organisms. Effective transduction with lentiviral pseudovirus particles does not require cells to be active or growing. The efficiency of lentiviral transduction is close to 100% so this system is ideal as an expression library vector.

Lentiviral Transcriptional Reporter Systems:

Eukaryotic gene expression is regulated by a wide variety of developmental and environmental stimuli. First, an extracellular signaling molecule binds to a specific receptor. The signal is then transmitted through a series of molecular cascades, which activate or deactivate specific transcription factors (TFs) that regulate gene expression. The expression of any given gene is controlled by multiple transcription factors, which in turn are modulated by multiple signal transduction pathways. Many of these signal transduction pathways converge at transcription factors that bind to specific transcriptional response elements (TREs) found in the promoters of various genes and modulate the transcription of these genes. The activation of a signal transduction pathway (e.g. by growth factors, drugs, etc.) can therefore be monitored by the expression level of the reporter gene controlled by a promoter containing these response elements.

Instant and Accurate High-throughput Cloning into Lentivectors:
 



Features and Benefits of the Clone-it™ Lentivector Systems:

-
Ligase, recombinase, topoisomerase and restriction enzyme-free system
- Rapid and Directional cloning with no recombination errors
- Near 100% cloning efficiency
- Short hands-on time and adaptable for high-throughput cloning
- Lentiviral-based expression systems containing HIV backbone for high viral titers
- Reliable delivery to dividing or non-dividing cells
- Convenient sorting of transfected or transduced cells
- Create stable cell lines

 


 

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