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.
|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：
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 |
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 ||EGFP |
|YSH-AV002-shRNA ||mCherry |