Lentivirus Packaging

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Lentivirus Packaging

·    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 Lentivirus Adenovirus AAV
Origin HIV-1 Adenovirus type 5 AAV2
Genome Two linear and continuous single-stranded RNAs A linear and continuous double-stranded DNA A linear and continuous single-stranded DNA
Genome size ~9.2 kb (WT) ~36 kb (WT) ~4.7 kb (WT)
Expression Duration long-term stable expression Transient expression Long-term expression
Integrate to cell genome Yes No Low frequency
Immunogenicity Low High Very 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
·    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    ·
Standard Titer Turnaround Application
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:

Plasmid construction (Plasmid restriction enzyme digestion and sequencing report) Lentivirus packaging QPCR confirm titer
FAQ about lentivirus, click to show
Knockout lentivirus
·    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 series Vector Reporter gene; Selection marker
Lentiviral YKO plasmid YKO-LV001-single-gRNA EGFPv/mCherry,Puro/Neo
YKO-LV001-dual-gRNA EGFPv/mCherry,Puro/Neo
Lentiviral Cas9 plasmid YCas-LV001 EGFPv
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
·    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 series Vector Reporter 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
·    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 series Vector Reporter 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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