AAV Vector Design & Cloning

Adeno-associated virus (AAV) vectors are widely used in gene delivery and gene therapy due to their efficiency and safety. The first step in successful AAV vector-based research is designing a plasmid that can be effectively packaged into an AAV capsid. This process involves optimizing transgene expression strategies, selecting appropriate tags, managing AAV genome sizes, and choosing the right AAV plasmid backbones to ensure high transgene expression, efficient packaging, and targeted delivery of the AAV vector.

At Biohippo, we offer one-stop services for designing, constructing, and cloning customized AAV vectors, as well as downstream virus packaging service. Our team of molecular and AAV experts provides comprehensive support throughout the service process to meet your specific needs.

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Service Details & Workflow

1.   Full Support on AAV Vector Design

• Design your gene of interest (GOI): Provide us with your Gene of Interest (GOI) and its sequence, or collaborate with our team of experts to design a vector tailored to your experimental requirements. We offer one-stop services for a wide range of vectors, including those for overexpression, CRISPR gene editing (sgRNA), shRNA, miRNA, and more. Additionally, we provide sgRNA and shRNA selection and validation services to ensure you achieve optimal knockdown or knockout efficiency.

• Choose promoter(s) and regulatory elements: Usually the promoter sequence is immediately upstream of the coding sequence and controls the binding of RNA polymerase and trnascription factors. Therefore, promoters play an important role in determining where and when your gene of interest will be expressed.

• Tags, Fluorescence Reporters, and Linkers:  Adding tags in-frame to N-terminal or C-terminal of a native protein is a well-established strategy for many applications, including protein purification, IP, WB, and in vivo imaging. Some common tags are His, FLAG, HA, c-Myc, etc. Fluorescent proteins, such as GFP,  mCherry, tdTomato, are most often used for live cell imaging and FACS. Fluorescent protein gene can be directly fused to target genes or linked with 2A self-cleaving peptides (such as T2A and P2A). Moreover, Fluorescent protein can also be introduced by IRES or even by another promoter. 

• AAV Serotype: We offer a multitude of AAV serotypes, encompassing widely recognized AAV serotypes (AAV1-13), as well as engineered AAV capsids. Diverse selection of AAV serotypes enables distinct tropisms tailor for a wide range of applications.

2.   Synthesis of DNA fragments

• Design your gene of interest (GOI).
​• Synthesis of DNA fragments with low cost ($0.25/bp).
• Amplification of requested sequences.

3.    Molecular Cloning

• Cloning with standard cloning techniques, Gibson assembly or Gateway cloning. 
• Multiple fragments can be assembled at once.​

4.    Strict Quality Control

• Restriction enzyme digestion to identify correct clones.​
• SmaI/AdhI digestion to confirm two intact ITRs.
• Sanger sequencing to verify inserts.
• Whole plasmid sequencing available upon request.​​

5.    Delivery

• The experimental strategy, raw data and final results are summarized in a detailed report.
• The plasmid DNA (mini scale preparation) is delivered to customer. Higher scales available upon request.

​6.    Proceed to AAV packaging service, if applicable

• We offer AAV packaging service to streamline your workflow. Please click here for more details.

Other Viruses at Biohippo

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Biohippo offers a variety of viral vectors, including RV, HSV, PRV, and VSV, to replace conventional tracers with the highly efficient, more specific, and less invasive viral method.

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