Adeno-associated Virus Packaging

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Adeno-associated Virus PackagingService

·    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.

AAV Packaging Technical Detail
·    AAV Packaging Technical Detail    ·
Adeno-associated virus (AAV) is a kind of Parvovirus, which genome is single-stranded DNA. It can infect both divisive and non-divisive cells. Adenoviruses or herpes viruses are usually needed to assist AVV to replicate in vivo. Recombinant AAV (rAAV) is a viral vector that combines the AAV2 genome with capsid protein genomes of different serotypes. CDS or RNAi interference sequences of the target genes can be inserted into the rAAV plasmid. AAV is mainly used in vivo injection because it has low immunogenicity and long-term expression.
Ubigene’s AAV will be purified by ultracentrifugation, and its genome copy will be titrated by qPCR. The titer of AAV ranged from 10^12 to 10^13 v. g. / ml.

· Different serotypes of AAVs have different transduction efficiency for tissues and cells. There are 10 different AAV serotypes in our packaging service. It is highly recommended that customers should select the corresponding AAV serotypes for different tissues:

Serotype Tissues
AAV1 SM,CNS,lung,retina,pancreas,heart,liver
AAV2 SM,SNS,liver,kidney,retina
AAV4 CNS,retina,lung,kidney
AAV5 SM,CNS,lung,retina
AAV6 SM,heart,lung,adipose,liver
AAV7 SM,retina,CNS,liver
AAV8 SM,CNS,retina,liver,pancreas,heart,kidney,adipose
AAV9 SM,lung,liver,heart,pancreas,CNS,retina,testes,kidney
AAVrh10 SM,lung,liver,heart,pancreas,CNS,retina,kidney
AAVPHP.B CNS
AAVDJ/8 SM, liver, CNS, retina
AAVPHP.S PNS
AAVPHP.eB CNS(stronger than AAVPHP.B)

Product Specs:

Specs Titer Turnaround Application
Standard Size 1*10^11 GC 2-3weeks Cell transduction
Ultra-purified large size 1*10^12 GC 2-3weeks In vivo injection

· Work Flow and Validation:

Plasmid construction (Plasmid restriction enzyme digestion and sequencing report)

Lentivirus packaging

QPCR confirm titer

Knockout AAV
·    Gene Knockout AAV    ·
The gene knockout AAV vector would be constructed and packaged as AAV. After ultracentrifugation purification, it can be used for in vivo injection. Cells infected with AAV can stably express gRNA and Cas9 protein to knockout the target gene.

· Gene knockout of AAV vector selection

YKO series Vector Reporter gene; Selection marker
AAV YKO plasmid YKO-AAV001-gRNA EGFPv
YKO-AAV002-gRNA mCherry
Due to the limited capacity of AAV, SaCas9 protein can be used. SaCas9 and gRNA on the same construct can increase transduction efficiency.
Expression AAV
·    Gene expression AAV    ·
The gene overexpression vector would be constructed and packaged as AAV. After ultracentrifugation purification, it can be used for in vivo injection.
YOE series Vector Reporter gene; Selection marker
AAV YOE plasmid YOE-AAV001 EGFP
YOE-AAV002 mCherry
Customers can choose a variety of tissue-specific promoters that can achieve tissue-specific expression in vivo.
Customers can choose a variety of tissue-specific promoters that can achieve tissue-specific expression in vivo,Click to show
Knockdown AAV
·    Knockdown AAV    ·
The gene knockdown vector would be constructed and packaged as AAV. After ultracentrifugation purification, it can be used for in vivo injection.
YSH series Vector Reporter gene; Selection marker
AAV YSH plasmid YSH-AAV001-shRNA EGFPv
YSH-AAV002-shRNA mCherry
Customers can choose a variety of tissue-specific promoters that can achieve tissue-specific expression in vivo.


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