Applications: High Titer Virus Production
TransIT®-Lenti for High Titer Lentivirus Production
TransIT®-Lenti Transfection Reagent is designed to enhance delivery of packaging and transfer vectors to adherent HEK 293T and suspension 293-F cell types to increase recombinant lentivirus production.
- Provide up to eight-fold higher functional titers
- Simple protocol - no media change required, single harvest
- Animal origin free formulation
NEW Reference Card: Methods to Measure Lentivirus and AAV in Your Sample
Glossary of Lentivirus Terms Click to expand
Term |
Definition |
Functional Titer |
A measure of the amount of infectious virus particles (e.g. containing envelope, capsid and genome). Functional titer is often reported in Transducing Units per milliliter (TU/ml). |
LTR |
Long Terminal Repeats are regions of the RNA genome associated with regulation, integration and expression of retroviruses. In a lentiviral transfer plasmid, LTR sequences flank the gene of interest and aid in generating viral dsDNA by reverse transcription and integration into the host genome. |
MOI |
Multiplicity of Infection refers to the ratio of the number of virus particles to the number of target cells. |
Packaging Plasmids |
Packaging plasmids include the essential structural (gag), enzyme (pol), packaging (rev) and envelope (VSV-G) components of the lentivirus genome. For 2nd generation packaging plasmids, gag, pol and rev are encoded on a single vector, whereas VSV-G is on a separate vector. For increase safety, 3rd generation vectors separate gag and pol from rev in addition to maintaining VSV-G on a separate vector. Third generation vectors often have additional safety parameters such as fewer wild-type HIV sequences and self-inactivating (SIN) elements. |
P24 Protein |
P24 is a protein component of the lentivirus capsid. Assays that detect P24 protein are commonly used to quantify lentivirus. However, these assays overestimate functional virus titers because they detect defective virus particles and free P24 protein in addition to competent particles. For lentivirus, 100-1000 defective virus particles are produced for every functional virus particle. |
Transduction |
The process by which foreign DNA is introduced to the cell by a virus. |
Transduction Reagent |
A chemical compound that increases virus aggregation and uptake into a target cell type. |
Transfection |
Transfection is the process by which molecules such as DNA, RNA, oligonucleotides and protein are delivered to cells by non-viral (e.g. chemical or mechanical) methods. In the context of recombinant virus production, transfection is used to deliver plasmid DNA encoding the essential virus components (packaging plasmids) and the gene of interest (transfer plasmid) to producer cells such as HEK 293T/17. |
Transfer Plasmids |
The transfer plasmid encodes the gene of interest flanked by long terminal repeat (LTR) sequences, as well as other essential components such as RNA packaging signal (Psi) and rev response element (RRE). |
VSVG |
The Vesicular Stomatitis Virus G glycoprotein is a broad tropism envelope protein that is frequently used to pseudotype recombinant lentivirus. VSV-G also increases lentivirus stability in downstream purification processes. |
Technical Resources
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