商品编号


MIR 6603



品牌


Mirusbio



公司


Mirus Bio



公司分类


Transfection



Size

0.3 ml

商品信息


Product Overview


Trans
IT?-Lenti Transfection Reagent is designed to enhance delivery of packaging and transfer vectors to adherent HEK 293T and s
USP
ension 293-F cell types to increase recombinant lentivirus production.


High Performance
?
–?
Provide up to eight-fold higher functional titers

Simple Protocol

–?
No media change required, single harvest

Animal Origin Free?
–
?
Regulatory friendly


?
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

L
ong
T
erminal
R
epeats 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

M
ultiplicity
o
f
I
nfection 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.



Transdcution

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
V
esicular
S
tomatitis
V
irus
G
glycoprotein is a broad tropism envelope protein that is frequently used to pseudotype recombinant lentivirus. VSV-G also increases lentivirus st
ABI
lity in downstream purification processes.





Overview of Recombinant Lentivirus Production
Click to expand




Overview of Recombinant Lentivirus Particles and Infection of Target Cell.
(1) Producer cells (e.g. 293T) are transfected with 3-4 plasmids encoding the gene of interest, vesicular stomatitis G protein (VSV-G) and essential virus proteins (e.g. gag, pol and rev). (2) Virus is assembled and released into the supernatant through budding with the producer cell plasma membrane resulting in an envelope decorated with VSV-G. The medium containing virus is filtered through a 0.45 ?m filter to remove any cells. (3) Target cells are frequently transduced with recombinant lentivirus particles in the presence of a polycation to increase aggregation and uptake. The virus enters the cell and the capsid is uncoated revealing the RNA genome and viral enzymes. The viral RNA is reverse transcribed into DNA which is then integrated into the host genome. (4) Transcription and translation result in the production of the protein encoded by the gene of interest.



?

Specifications
Storage Conditions:
All Configurations:
Store at -20°C
Product Guarantee:
Trans
IT?-Lenti Transfection Reagent:
1 year
Transduce
IT? Transduction Reagent:
6 months