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Viral Vector Design and Development

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The choice of gene delivery tools is the key to successful gene therapy. Whether the drug can ultimately exert efficacy in the human body depends to a large extent on a stable and efficient delivery system. At present, the commonly used vectors for gene therapy include viral vectors and non-viral vectors. Viral vectors have the advantages of a wide host range, long acting time, and they can perform overexpression, interference, knockout, endogenous activation of coding or non-coding genes, so they are more and more widely used in gene therapy research. Adeno-associated virus (AAV), adenovirus (ADV) and lentivirus (LV) are all commonly used viral vectors with different infection characteristics. Among them, AAV is currently the most widely used delivery vector in gene transduction and gene therapy due to its advantages of good tissue specificity, low immunogenicity, and high safety.

Cyagen has many years of experience in virus packaging platforms, and can provide AAV, LV, ADV and other viruses that comply with different quality standards. Our virus packaging platform has been widely used in the construction of various cell line models and in vivo studies of living animals, and have been cited and published by customers in many documents. For AAV virus packaging, we adopt a three-plasmid co-transfection method. The advanced purification process can provide customized AAV packaging services with high purity, high titer, and different serotypes. It is especially suitable for in vivo animal experiments and can meet the personalized choices of gene therapy researchers.
Cyagen can provide the following AAV packaging services (and more upon request):

AAV Type

Vector Production Scale

Virus amount




Purified pilot

≥1*10^12 GC

≥1*10^12 GC/mL

5~6 weeks

Purified medium

≥5*10^12 GC

≥1*10^13 GC/mL


Purified pilot(3+1)

≥1*10^12 GC

≥1*10^12 GC/mL

Purified medium(3+1)

≥5*10^12 GC

≥1*10^13 GC/mL


Purified pilot

≥1*10^12 GC

≥1*10^12 GC/mL

Purified medium

≥5*10^12 GC

≥1*10^13 GC/mL

Note: Conventional projects will receive a complimentary control virus.
Cyagen can provide the following AAV serotypes (and more upon request):


Target tissue


Target tissue


muscles, heart, nerves, skeletal muscles

AAV rh10

liver, blood, heart, extracorporeal cells


muscles, liver, brain tissue, eyes, nerves


nerve cell (reverse non-transsynaptic)


lungs, eyes, nerves, pancreas


retina, lung, kidney, cells in vitro


mungs, heart


liver, eyes, central nervous system


muscle, liver


central nervous system


muscles, liver, nerves, eyes


peripheral nervous system


heart, muscles, lungs, liver, nerves

More serotypes

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Advantages of Cyagen’s AAV virus packaging service:

  • Short cycle: Virus packaging completed in as fast as 2 weeks, and customized projects can be as fast as 5 weeks (additional time for delivery).
  • Reliable quality: The virus titer is guaranteed to be ≥1*10^12 GC/mL, the purity is ≥95%, and the endotoxin is less than 10 EU/mL, ensuring high infection efficiency.
  • Professional support: We provide professional technical guidance from vector design, virus packaging and use, in vivo injection to phenotypic analysis.
  • Rich experience: More than ten years of virus packaging experience, continuously expanding stock library, and numerous customer literature citations and publications.
  • Artificial Intelligence (AI) Bioinformatics Platform: Our experienced artificial intelligence (AI) and bioinformatics team provides cutting-edge AAV carrier development, optimization, and transformation services.
AAV virus packaging technology route:

In addition to AAV virus vector packaging, Cyagen can provide the following LV and ADV virus packaging:




Lentivirus (LV)


≥1*10^8 TU/mL

5-8 weeks


≥1*10^9 TU/mL

Adenovirus (ADV)


≥1*10^10 PFU/mL

7-10 weeks


≥1*10^12 PFU/mL

Frequently asked questions about virus packaging:

Q1: What is the difference between AAV, LV, and ADV viral vectors?






Particle diameter





4.7kb single-stranded DNA or 2.4kb double-stranded DNA

36 kb double-stranded DNA

9.3 kb double-stranded RNA






Phased expression at medium to

high level

High-level transient expression

Moderately stable expression

Expression Duration

Begin to express within 7-14 days,

Begin to express in 1-2 days,

Begin to express in 2-4 days,

Generally 6 months to 24 months

Lasts ≤ (2-4) weeks

Lasts ≥6 months

Diffusion capacity in vivo





Very low



Integration method

Small amount of targeted integration

No integration

High frequency random



High yield, simple composition, good safety

High yield and easy purification

Low yield, stable expression


Animal experiments

Cell/animal larger gene


Cell experiments


Q2: Why is AAV the most popular vector in the field of gene therapy?

​​Because AAV has the following advantages:

​​1. Wide host range: dividing and non-dividing cells;

2. High safety: No pathogenic AAV is found; ITR and rep/cap are expressed on independent plasmids;

3. Low immunogenicity: The immune response to AAV in animals or humans is the smallest among several viral vectors;

4. Good targeting: a variety of serotypes and tissue specificity options.

Q3: What are the key points that need to be paid attention to when implementing AAV expression in animals?

1. Selection of serotype. The serotype of AAV is mainly determined by the structure of the AAV capsid protein. Different capsid protein structures recognize different cell surface receptors. Therefore, the choice of serotype will affect the infection efficiency, tissue affinity, and start time of AAV expression.

2. The choice of promoter. Broad-spectrum promoter or specific promoter? Compared with the broad tissue specificity of serotypes, specific promoters can achieve specificity by cell type. Therefore, for studies with higher specificity requirements, a certain cell-specific promoter can be selected.

3. Injection method. For tissues and organs that can be injected locally, local multipoint injection of target organs can achieve better specificity and expression effects.

4. Virus amount and titer. The recommended amount of AAV injected differs according to the target organs/tissues and AAV serotypes used (different diffusion capacity), and the amount of virus particles delivered to the tissue has a great influence on the infection effect. However, the amount of AAV used is not as much as possible — too much virus may sometimes even reduce expression. If you want to know the best viral administration process for a specific application target, you can contact us for consultation.

5. Detection time. The peak time of expression of different genes is different, and AAV needs to change from single-stranded DNA to double-stranded DNA. Gene expression can generally be detected within 2 weeks. If there is no antibody impact, the modified gene can continue to be expressed for more than half a year.

Q4: How much virus is sufficient for AAV injection in mice?

There are many factors that affect the total amount of AAV injection. The most important ones are serotype, target tissue type, injection method (local or systemic), and animal model (such as rat or mouse). For those reported in the literature, please refer to the recorded injection methods for your application; it is considered best practice to set several gradients in initial experiments. For example, using AAV9 to infect mouse heart tissue (cardiomyocytes), either tail vein injection or myocardial multi-point injection is recommended. Tail vein injection recommends 1011 vg/mL, 100 μL; origin injection is recommended 1010 vg/mL, 20 μL/point. If you want to know more about the AAV virus injection recommendations of different applications, please contact us.

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