Overview of AAV sequencing

Adeno-associated virus (AAV) is the most commonly used gene editing vector in preclinical and clinical studies. AAV is an envelope-free linear double-stranded DNA virus that is widely distributed in nature and exists in at least 100 serotypes. Its genome is approximately 36 kb long with an inverted terminal repeat region (ITR) at each end and a viral packaging signal on the inner side of the ITR. The ITR is lined with viral packaging signals. Four early transcripts (E1, E2, E3, E4) are distributed on the genome and are responsible for regulatory functions, and one late transcript is responsible for encoding structural proteins.

Figure 1. Encapsulated content of AAV virus.

CD Genomics has been providing you with one-stop NGS-based AAV-related services, which will greatly simplify the process of AAV development and evaluation. Our professional team of experts, standardized AAV sequencing QC process and best-in-class sequencing technology platform ensure that we can assist you in achieving more in-depth and complex applied research in AAV development.

The workflow of AAV sequencing service

CD Genomics' goal is always to provide the highest quality NGS-based standardized AAV sequencing to be your hands-on assistant in AAV research. Our AAV sequencing services and solutions are focused on these areas, such as AAV whole genome sequencing, AAV-ITR sequencing, quantification of AAV expression and host response assay analysis. The main processes of AAV sequencing are shown below, including AAV sample preparation, AAV library preparation, up-sequencing and bioinformatics analysis. Our sequencing process is governed by strict QC standards, ultimately ensuring the most reliable data analysis reports for you with the fastest turnaround time.

01. Sequencing sample requirements

02. Single cell library construction and NGS

NGS-based Illumina / MGI platform

03. Bioinformatics Analysis

• Alignment with plasmid sequences and host reference genome
• Checking genome integrity
• Heterogeneity detection
• AAV-ITR sequencing
• Confirmation of plasmid sequences
• Validate packaging materials, DNA contaminant identification
• Quantify AAV expression
• Host immune response for stratification and characterization

Advantages of our AAV sequencing service

• Our AAV sequencing service offers significant advantages in fast-read sequence confirmation of difficult ITR regions, accelerating the screening and validation process of lead candidates
• One-stop solution for efficient NGS-based AAV sequencing from sample to report
• The most stringent quality control, optimized, highly automated and scalable workflow for high quality, reproducible and reliable analysis results
• Optional NGS-based standardized and customizable solutions

Applications

The NGS-based AAV sequencing service provided by CD Genomics is of great relevance to research projects on AAV vectors. First, high quality and efficient AAV sequencing services are mainly applied to the process of designing, improving and modifying AAV vectors. In addition, characterization, characterization and quantification of AAV can be achieved. Finally, the AAV sequencing results allow us to determine the safety and efficacy of AAV vector system.

CD Genomics is committed to combining professional project protocol guidance and standardized analysis processes to ensure that your project is performed accurately and quickly. If you are interested in CD Genomics' AAV sequencing service, please feel free to contact us now.