Adeno-Associated Virus (AAV), one of the most popular vectors in gene therapy, is renowned for its low pathogenicity, long-term expression, and favorable safety profile. However, upon entry into host cells, the AAV vector genome has the potential to randomly integrate into host chromosomes. While this incidence is much lower than with other retroviral vectors, this integration event carries potential risks, such as insertional mutagenesis activating oncogenes or inactivating tumor suppressor genes. Therefore, accurate detection of AAV integration sites is crucial for assessing the long-term safety of gene therapy products, optimizing vector design, and advancing clinical research.

AAV Integration Site Analysis is a technical service that utilizes high-throughput sequencing (NGS) and bioinformatics analysis to comprehensively and accurately identify the insertion location and frequency of the AAV vector genome within the host cell chromosome, as well as its surrounding genomic context. Its core purpose is to assess the integration potential and potential genotoxicity risks of AAV vectors in gene therapy products, providing critical safety data for preclinical and clinical trials.

AAV vectors primarily exist within cells as circular episomes (episomes), but a small portion integrates into chromosomes through host cell DNA repair mechanisms (such as non-homologous end joining (NHEJ) or microhomology-mediated end joining (MMEJ).

Currently, the mainstream detection technology is based on next-generation sequencing (NGS). Its core principles are:
1. Bridging and Enrichment: Utilizing the unique structure at the junction of the host genome and AAV vector sequences for enrichment. The most common methods are ligation-mediated PCR (LM-PCR) or linear amplification-mediated PCR (LAM-PCR). These methods use specific primers to selectively amplify fragments containing only the "host-DNA-AAV vector-DNA" junction, significantly enriching for integration site information while eliminating interference from large amounts of background genomic and episomal DNA.

A complete AAV integration site analysis service typically includes the following steps:

LM-PCR/NGS-based technology can detect rare integration events with frequencies below 0.0001%, far exceeding the detection limit of traditional methods.

Integration sites can be discovered genome-wide, eliminating the need to pre-emptively assume integration hotspots.

The data and report formats provided comply with the FDA and other regulatory agency guidelines for the safety evaluation of gene therapy products, providing solid data support for IND filings.

Our experienced team handles complex experimental and data analysis processes, significantly saving R&D personnel time and effort and accelerating the development process.

We not only provide a list of integration sites but also provide in-depth interpretation based on genomic biology, providing clients with actionable insights.