| Choose your Viral Vector for Gene Delivery |
| |
Adenovirus |
Adeno-Associated Virus (AAV) |
Lentivirus (HIV-1, FIV, SIV) |
Retrovirus
(MMLV) |
| Gene Expression |
Transient |
Transient or Stable |
Transient or Stable |
Stable |
| Infect Dividing Cells |
Yes |
Yes |
Yes |
Yes |
| Infect Non-dividing Cells |
Yes |
Yes |
Yes |
No |
| Integration into Target Cell Genome |
No |
Yes |
Yes |
Yes |
| Immune Response in Target Cells |
High |
Very Low |
Low |
Moderate |
| Relative Viral Titer |
XXXX |
XXX |
XXX |
XX |
| Relative Transduction Efficiency |
XXXX |
XXX |
XXX |
XX |
AAV Expression Procedure
1. Package Virus with Gene of Interest |
2. Measure Viral Titer  |
3. Purify your AAV
 |
4. Infect Target Cells |
AAV Helper Free Expression and Packaging Systems
Production of recombinant adeno-associated virus requires certain genes from the adenovirus genome, which means that a “helper” adenovirus usually needs to be present during viral packaging. Our AAV Helper Free Expression Systems eliminate the need for a helper adenovirus. Most of the required adenoviral genes (E2A, E4 and VA RNA) are provided in a pHelper plasmid, while the E1 gene is provided by the 293 cells. We offer the most comprehensive and flexible line of AAV Helper Free Expression Systems, Packaging Systems, and Expression Vectors commercially available. Choose from 6 native AAV serotypes or our new DJ and DJ/8 systems (see details on following page). In addition, you can buy only the plasmids you need:
- Complete Expression Systems include packaging vectors, an expression vector to clone your gene of interest, and a GFP control vector
- Packaging Systems without the expression vector
- Expression Vectors alone (choice of 6)
When packaging your adeno-associated virus, consider our 293AAV Cell Line. This cell line is derived from the parental HEK 293 cell line, but has been specifically selected for use in AAV expression. You’ll see firmer attachment to culture plates, flattened morphology, and a much larger surface area for higher viral yields and superior transfection.
Select your AAV Helper Free Vectors
First determine which plasmids you need to construct your virus, then use the table below.
| |
AAV Helper Free
Expression Systems |
AAV Helper Free
Packaging Systems |
AAV Helper Free
Expression Vectors |
| Expression Vector |
X |
|
X |
| pHelper Vector |
X |
X |
|
| pRC Vector |
X |
X |
|
| GFP Control Vector |
X |
X |
|
| Application |
Best value: all the plasmids you need to make a recombinant virus containing your gene of interest |
Co-transfect pHelper and pRC plasmids with your own AAV expression vector |
Clone your gene of interest here and co-transfect with an AAV packaging system |
Select your AAV Serotype
When selecting an AAV Helper Free Expression System or Packaging System, choose from 8 serotypes:
- Native serotypes AAV-1 through AAV-6
- AAV-DJ, engineered by DNA family shuffling to create a hybrid capsid from 8 native serotypes; AAV-DJ vectors convey significantly higher in vitro infectivity rates compared to native serotypes across a broad range of cells and tissues (see table below)
- AAV-DJ/8, a mutant of AAV-DJ that exhibits increased uptake in brain and other tissues in vivo, similar to native serotypes 8 and 9
Relative In Vitro Infectivity Rates of AAV Serotypes*
| Cell Line |
AAV-1 |
AAV-2 |
AAV-3 |
AAV-4 |
AAV-5 |
AAV-6 |
AAV-8 |
AAV-9 |
AAV-DJ |
AAV-DJ/8 |
| Huh-7 |
13 |
100 |
2.5 |
0.0 |
0.1 |
10 |
0.7 |
0.0 |
500 |
0.2 |
| HEK293 |
25 |
100 |
2.5 |
0.1 |
0.1 |
5 |
0.7 |
0.1 |
500 |
0.3 |
| HeLa |
3 |
100 |
2.0 |
0.1 |
6.74 |
1 |
0.2 |
0.1 |
667 |
0.2 |
| HepG2 |
3 |
100 |
16.7 |
0.3 |
1.7 |
5 |
0.3 |
ND |
1250 |
0.5 |
| Hep1A |
20 |
100 |
0.2 |
1.0 |
0.1 |
1 |
0.2 |
0.0 |
400 |
0.1 |
| 911 |
17 |
100 |
11 |
0.2 |
0.1 |
17 |
0.1 |
ND |
500 |
0.0 |
| CHO |
100 |
100 |
14 |
1.4 |
333 |
50 |
10 |
1.0 |
25000 |
5.0 |
| COS |
33 |
100 |
33 |
3.3 |
5.0 |
14 |
2.0 |
0.5 |
500 |
0.3 |
| MeWo |
10 |
100 |
20 |
0.3 |
6.7 |
10 |
1.0 |
0.2 |
2857 |
1.0 |
| NIH3T3 |
10 |
100 |
2.9 |
2.9 |
0.3 |
10 |
0.3 |
ND |
500 |
0.1 |
*Normalized to AAV-2 = 100. ND = Not determined.
Once you’ve harvested your AAV you will want to measure the titer of your virus prior to infecting your target cells. We have developed a quick, easy method to quantify the AAV particles in your viral stock. The QuickTiter™ AAV Quantitation Kit measures the concentration of AAV in genome copies per mL (GC/mL) in less than two hours. Simply prepare a standard curve using the DNA standard included.
The standards are run alongside your samples in any fluorescence plate reader. Compare the relative fluorescence units (RFU) of your sample to the standard curve to determine your AAV titer. The QuickTiter™ AAV Quantitation Kit will quantify purified AAV of any serotype. If you decide not to purify your AAV, the QuickTiter™ AAV Quantitation Kit works just as well on unpurified AAV-2 and AAV-DJ.
Purification of your AAV is important in obtaining good results, but the thought of time
-consuming ultracentrifugation might tempt you to skip this crucial step. ViraBind™ AAV Purification Kits use a one-step proprietary matrix followed by further purification and concentration using a centrifugal concentrator. The result is a higher AAV yield with higher purity in a fraction of the time. ViraBind™ AAV Purification Kits are available in two sizes:
- The standard kit can process 1 x 15cm dish or 2 x 10cm dishes per prep
- The mega kit can process 10 x 15cm dishes per prep