Adenovirus Packaging

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Adenovirus 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.

Adenovirus Packaging Technical Details
·    Adenovirus Packaging Technical Details    ·
Adenovirus is a linear and double-stranded DNA virus that without a surface envelope. It can infect both divisive and non-divisive cells. Recombinant adenovirus can enter cells through receptor-mediated endocytosis, while its genome is not integrated with the host cell genome. Adenovirus is widely used in the researches of cardiovascular, liver, muscle, lung, cancer and other fields. It has the characteristics of wide host range, strong immunogenicity, large capacity, no integration with the genome, transient expression and so on. To ensure the safety, human adenovirus type 5 (Ad5), which is a replication-incompetent (E1 and E3 genes deleted) adenovirus, is used. E1 is essential for the replication of adenovirus. The absence of E1 prevents its self-replication. Its replication depends on packaging cells such as 293A cells, which provide trans-complementary replication for adenovirus. The protein expressed by the E3 gene can resist the host's antiviral defense system, and the removal of the E3 region can reduce the host's immune response. The adenovirus particles provided by Ubigene would be centrifuged and filtered by gradient ultracentrifugation, and the titer would be determined by chitin immunoassay. The titer of adenovirus is in the range of 10 ^ 10 ~ 10 ^ 12pfu / ml.
·    Product Specs    ·
Titer Turnaround Application
Standard Size 1*10^10 PFU 3-4weeks Cell transduction
Large size 1*10^11 PFU 3-4weeks Cell transduction
Ultra-purified 1*10^10 PFU 3-4weeks In vivo injection

· Work Flow and Validation:

Plasmid construction (Plasmid restriction enzyme digestion and sequencing report) Lentivirus packaging Titer confirmation
Gene expression adenovirus
·    Gene expression adenovirus    ·
The gene expression plasmid would be constructed and packaged as adenovirus. After infecting host cells with overexpression adenovirus, host cells would transiently overexpress the target gene. The capacity of adenovirus is as large as 4.5kb.

· Gene knockout of Adenovirus vector selection

YOE series Vector Reporter gene; Selection marker
Adenoviral YOE plasmid YOE-AV001 EGFP
YOE-AV002 mCherry
Gene knock of Adenovirus vector selection
·    Gene knockdown adenovirus    ·
The shRNA expression vector would be constructed and packaged as adenovirus. After infecting host cells with adenovirus, host cells would transiently express shRNA, resulting in the RNA interference-effect.

· Gene knockdown of Adenovirus vector selection :

YSH series Vector Reporter gene; Selection marker
Adenoviral YSH plasmid YSH-AV001-shRNA EGFPv
YSH-AV002-shRNA mCherry

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