ORIGENE에 제공하는 고품질의 AAV packaging service를 이용해 보세요 !
Adeno-Associated Virus (AAV)는 단일 가닥의 DNA를 갖고 있는 non-enveloped virus의 일종으로 분열/비분열 세포 모두에 유전자 전달과 발현이 가능하다는 장점을 가지고 있습니다. Recombinant AAV (rAAV)는 target gene의 전달 효율을 높일 수 있기 때문에 특정 세포주에 관심 있는 유전체를 전달하는 연구용 도구로 많이 사용하고 있습니다.
Virus |
Expression |
Genome |
Packaging Capacity |
Virus Size (nm) |
Cells Infected |
Target Cell Genome Integration |
Immune Response |
Lentivirus |
Stable |
RNA |
< 8kb |
80-130 |
Dividing/Non-dividing |
Yes |
Low |
AAV |
Transient or Stable* |
ssDNA (linear) |
~4.5kb |
18-26 |
Dividing/Non-dividing |
No* |
Very Low |
Adenovirus |
Transient |
dsDNA (linear) |
>8kb |
105 |
Dividing/Non-dividing |
No |
High |
Y-Retrovirus |
Stable |
RNA |
<8kb |
80-130 |
Dividing |
Yes |
Moderate |
*Recombinant AAV has a low frequency of target cell genome integration
Recombinant AAV Packaging Workflow
In vitro transduction efficiency of HEK293T cell line with different AAV serotypes.
- 11 different serotypes available: AAV1, AAV2, AAV3, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh.10, AAVDJ, AAV-PHP.eb
- Fast turnaround time: 2-3 weeks
- Competitive titer: > 1×1013 GC/mL guaranteed
- Flexibility: Customizable volumes, titers, and purification
AAV Expression Vectors
Code | Description |
---|---|
PS100089 | pAAV-AC-Myc-DDK, AAV vector with C-terminal Myc-DDK tag |
PS100090 | pAAV-AC-GFP, AAV vector with C-terminal tGFP tag |
PS100091 | pAAV-AC-RFP, AAV vector with C-terminal tRFP tag |
PS100127 | pAAV-EF1a-tGFP-WPRE AAV Gene Expression Vector |
TR30034 | HuSH shRNA GFP AAV Cloning Vector (pGFP-A-shAAV Vector), 5 ug |
Serotype Selection
Tissue | Recommended Serotypes |
---|---|
Muscle | AAV-1, AAV-6, AAV-7, AAV-8, AAV-9 |
Liver | AAV-7, AAV-8, AAV-9, AAV-DJ |
Lung | AAV-4, AAV-5, AAV-6, AAV-9 |
CNS | AAV-1, AAV-2, AAV-4, AAV-5, AAV-8, AAV-9,AAV-PHP.eB, AAV-rh10, AAV-DJ |
Retina | AAV-1, AAV-2, AAV-4, AAV-5, AAV-8 |
Pancreas | AAV-8 |
Kidney | AAV-2, AAV-9 |
Heart | AAV-1, AAV-8, AAV-9 |
Rajawat, Yogendra Singh et al. “In-Vivo Gene Therapy with Foamy Virus Vectors.” Viruses vol. 11,12 1091. 23 Nov. 2019, doi:10.3390/v11121091
Q&A :
What is the difference between AAV and Adenovirus?
There are several key distinctions between them, including:
1) Packaging capacity
2) Onset and duration of gene expression
3) Immune response
Adenoviruses have a capacity of ~8.5 kilobases, and onset of expression can occur as early as 16-24 hours after infection. The high immune response from the target cells is the main limitation of adenoviral systems. Despite this, they are still widely used in research, due to their highly efficient transduction of most tissues.
AAVs have a packaging capacity of ~4.5 kilobases and have a much later onset of expression (2-7 days for in vitro and 3-21 days for in vivo), however, this delivery system triggers very low levels of immune response.
I have transduced my cells with AAV particles, but I still do not see any gene expression. What could be the problem?
You may not have produced a high enough titer. This could be caused by the health of the HEK293/HEK293T cells. HEK293T cells usually lose packaging efficiency after many passages. Cells should not be used after culturing for 1-2 months. Cells are also happier when seeded the day before transfection, which results in higher transfection efficiency.
You may also want to consider revisiting the transfection step and ensuring that you avoid contamination, have an effective transfection reagent, and lastly, consider the size of your insert
How do you calculate the MOI for AAV?
To determine the MOI needed when transducing a cell line for the first time, we recommend transducing the target cells with the same serotype eGFP reporter AAV particles at a series of MOIs, such as MOIs of 100, 1,000, and 10,000, etc…
Example: 3 x 105 HT1080 cells were prepared for transduction by AAV particles (titer: 1 x 1011 GC/ml) one day after plating. For an MOI = 1,000, 3μl AAV particles are needed. For an MOI = 10,000, 30μl AAV particles are needed.
Would you recommend using alkaline electrophoresis for measuring the titer?
We recommend measuring the titer using qPCR.