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Overview

Hysigen provides lentivirus, adenovirus and adeno-associated virus (AAV) with different sizes to achieve knockout (KO), overexpression and knockdown in cell models easily.

Comparison of different viruses

Lentivirus Packaging Technical Information

The lentiviral vector is derived from HIV-1 (Human Immunodeficiency Virus-1) and constructed using the VSVG capsid from the herpes virus. Functioning as a pseudovirus, it eliminates pathogenic genes, substituting them with exogenous target genes. Lentiviruses exhibit a broad range of infections and proficiently infect both dividing and non-dividing cells. Lentiviruses facilitate the effective integration of exogenous genes into host chromosomes, ensuring continuous expression. Consequently, lentiviruses have evolved into valuable tools for introducing exogenous genes. They find extensive application in diverse cell lines for purposes such as gene knockin/knockout, overexpression, RNA interference, microRNA-related research, and in vivo experiments.

AAV Packaging Technical Information

Adeno-associated virus (AAV) belongs to the Parvovirus family, featuring a single-stranded DNA genome. It exhibits the capability to infect both dividing and non-dividing cells, often requiring the assistance of adenoviruses or herpes viruses for in vivo replication. Recombinant AAV (rAAV) serves as a viral vector, merging the AAV2 genome with capsid protein genomes from various serotypes. The rAAV plasmid allows the insertion of coding sequences (CDS) or RNA interference sequences targeting specific genes.

AAV is primarily administered through in vivo injection due to its low immunogenicity and ability to ensure long-term expression. Hysigen's AAV undergoes purification via ultracentrifugation, and its genome copy is quantified using quantitative polymerase chain reaction (qPCR). The AAV titer typically ranges from 10^12 to 10^13 viral genomes per milliliter (v.g./ml).

Adenovirus Packaging Technical Information

Adenovirus is a linear, double-stranded DNA virus lacking a surface envelope. It demonstrates the ability to infect both dividing and non-dividing cells. Recombinant adenovirus can enter cells through receptor-mediated endocytosis, and notably, its genome does not integrate with the host cell genome. Adenovirus finds extensive use in cardiovascular, liver, muscle, lung, cancer research, and various other fields due to its broad host range, strong immunogenicity, large capacity, lack of genome integration, and transient expression characteristics.

To ensure safety, a replication-incompetent adenovirus type 5 (Ad5) is employed, with the essential E1 and E3 genes deleted. E1 is crucial for adenovirus replication, and its absence prevents self-replication. Replication relies on packaging cells like 293A cells, providing trans-complementary replication for adenovirus. The E3 gene-expressed protein counters the host's antiviral defense system, and removing the E3 region diminishes the host's immune response.

Hysigen's adenovirus particles undergo centrifugation and filtration via gradient ultracentrifugation, with the titer determined through chitin immunoassay. The adenovirus titer typically falls within the range of 10^10 to 10^12 plaque-forming units per milliliter (pfu/ml).