Lentivirus Packaging

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Efficient transfection of viruses
Ubigene provides lentivirus, adenovirus and adeno-associated virus (AAV) with different sizes and grades of purification. Our products can easily achieve knockout (KO), overexpression and knockdown in vitro and in vivo.
Comparison of different viruses:
Type LentivirusAdenovirusAAV
Origin HIV-1Adenovirus type 5AAV2
Genome Two linear and continuous single-stranded RNAsA linear and continuous double-stranded DNAA linear and continuous single-stranded DNA
Genome size ~9.2 kb (WT)~36 kb (WT)~4.7 kb (WT)
Expression Duration long-term stable expressionTransient expressionLong-term expression
Integrate to cell genome YesNoLow frequency
Immunogenicity LowHighVery low
Capacity ~6.4 kb~8.3 kb~4.7 kb
Advantages

· Wide range of applications;
· High transduction efficiency;
· stable hereditary.

· High transduction efficiency;
· Large capacity;
· Expandable.

· Low immunogenicity;
· High safety;
· High specificity.

Lentivirus Packaging Technical Details
The lentivirus vector is based on HIV-1 (Human Immunodeficiency Virus-1) and developed from herpes virus VSVG capsid. It is a pseudovirus, as its pathogenic genes will be deleted and replaced by exogenous target genes. Lentiviruses have a wide spectrum of infections and can effectively infect divisive and non-divisive cells. Exogenous genes can be effectively integrated into host chromosomes by Lentiviruses, to achieve continuous expression. Thus, lentivirus has become a useful tool for introducing exogenous genes. Lentiviral systems have been widely used in various cell lines for gene knockin/knockout, overexpression, RNA interference, microRNA related researches and in vivo experiments.
Product Specs
StandardTiterTurnaroundApplication
Standard 1*10^8 TU 2-3weeks Cell transduction
Large size 1*10^9 TU 2-3weeks Cell transduction
Ultra-purified 1*10^9 TU 2-3weeks In vivo injection
Work Flow and Validation:
FAQ about lentivirus, click to show

·FAQ about lentivirus:

1. How long does it take for gene expression after transduction?
It takes a long time for expression after lentivirus transduction. But fluorescence can be observed 24 hours after transduction on the cells with strong metabolism (such as 293T, HEK293, etc.); the fluorescent protein expression of cells with slow metabolism (such as primary culture cells, neural stem cells, embryonic stem cells, etc.) takes a longer time, i.e. 72-96 hours or even longer after transduction. The transduced cells can be continuously cultured for one week, and the effect of transduction of the virus to the target cells can be determined by observing the expression time and intensity of fluorescence.
2. How to improve transduction efficiency?
Ensure that the cells are in good growth condition before transduction. Use 293T for titer test to verify whether the titer of virus drops significantly; properly increase the MOI and extend the transduction time.
3. After lentivirus transduction, the condition of cells is very poor or even a large number of deaths. How to solve this problem?
Poor cell conditions before transduction, exceeding MOI, and cytotoxicity of target genes may lead to abnormal cell conditions.The main solutions are:
(1) make sure the pre-transduction cell condition;
(2)reduce the MOI to verify the cytotoxicity of the target gene;
(3) observe the cells after 6 hours of transduction, change the fresh medium in case of abnormality of cells, if not, change the medium 24 hours later.


Knockout lentivirus
The lentivirus knockout plasmids would be packaged as lentiviruses.After the cells are infected with lentiviruses, it can stably express gRNA and Cas9 protein to knockout the target gene.
Knockout lentivirus vector selection:
YKO seriesVectorReporter gene; Selection marker
Lentiviral YKO plasmid YKO-LV001-single-gRNA EGFP/mCherry,Puro/Neo
YKO-LV001-dual-gRNA EGFP/mCherry,Puro/Neo
Lentiviral Cas9 plasmid YCas-LV001 EGFP
YCas-LV002 Hygro
Because of the limitation of lentivirus capacity, the Cas9 protein, which is ~4kb, should be packed separately from gRNA. Generally, the two lentiviruses are used to co-infect the host cells.
Expression lentivirus
The gene expression plasmid would be constructed and packaged as lentivirus. After infecting host cells with overexpression lentivirus, drug resistance carried by virus vector can be used for drug screening, so as to obtain stable cell lines expressing target genes. Lentivirus can be used in vitro (including cell lines, immortalized cells, primary cells, and stem cells) and in vivo.
Gene overexpression lentivirus vector selection:
YOE seriesVectorReporter gene; Selection marker
Lentiviral YOE plasmid YOE-LV001 EGFP/Puro
YOE-LV002 mCherry/Puro
YOE-LV003 EGFP/Neo
YOE-LV004 mCherry/Neo
YOE-LV005 Puro
YOE-LV006 EGFP
YOE-LV007 mCherry
YOE-LV008 Luciferase/Puro
Knockdown lentivirus
The shRNA expression vector would be constructed and packaged as lentivirus. After infecting host cells with lentivirus, drug resistance carried by virus vector can be used for drug screening, so as to obtain stable cell lines expressing shRNA. Lentivirus can be used in vitro (including cell lines, immortalized cells, primary cells, and stem cells) and in vivo.
Gene interference lentivirus vector selection :
YSH seriesVectorReporter gene; Selection marker
Lentiviral YSH plasmid YSH-LV001-shRNA EGFP/Puro
YSH-LV002-shRNA mCherry/Puro
YSH-LV003-shRNA EGFP/Neo
YSH-LV005-shRNA EGFP/Hygro
YSH-LV004-shRNA mCherry/Neo
YSH-LV006-shRNA Puro
YSH-LV007-shRNA EGFP
YSH-LV008-shRNA mCherry
YSH-LV009-shRNA Luciferase/Puro

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