AVnerGene offers AAV capsid screening and evolution services through its ATHENA platform(ATHENA I and II). This platform allows for the high-throughput screening of AAV capsid libraries to identify the most efficient capsids for specific cell types or tissues. The ATHENA platform uses advanced techniques such as next-generation sequencing (NGS) to analyze the capsids and select the most promising candidates. AAVnerGene can also use directed evolution strategies to improve AAV capsid properties, such as tissue tropism and transduction efficiency, to better meet the needs of specific gene therapy applications.
AAVnerGene offers AAV capsid screening services for various animal model and cell lines, including mouse, non-human primate, and human cell lines and primary cells. This allows customers to select the best AAV capsid for their specific research or therapeutic needs, based on the target tissue or disease. The screening process can be customized based on the specific requirements of the customer’s project, and AAVnerGene’s experienced team can provide guidance and technical support throughout the process.
AAVnerGene has extensive experience in NGS library preparation and analysis. Our team is skilled in designing and constructing high-quality libraries for AAV capsid screening, as well as performing thorough analysis of the sequencing data. We use state-of-the-art sequencing technologies and bioinformatics tools to ensure accurate and reliable results. Our NGS library preparation and analysis services are available to customers who want to screen AAV capsids for their gene therapy applications.
Workflow of Capsid Selection and Evolution
The workflow for capsid evolution and screening in AAV typically involves several steps to generate and identify AAV capsids with desired properties. Here is a general overview of the process:
Library Generation: Start by creating a diverse library of AAV capsid variants. This can be done through various methods such as random peptide insertion(Such as ATHENA II), DNA shuffling(Such as ATHENA III), or other strategies(such as ATHENA I). The library should contain a large number of unique capsids with sequence diversity.
Packaging and Production: The AAV library is then packaged into viral particles using a packaging system(Such as mini-pHelper-Rep) that includes the necessary helper and replication components. This step involves co-transfection of the AAV library plasmids with the appropriate helper plasmids into host cells, which leads to the production of a viral library containing diverse AAV capsids.
Transduction and Selection: The viral library is then used to transduce target cells or tissues of interest. The selection process depends on the specific application and desired outcome. For example, if the goal is to identify capsids with enhanced tropism for a particular cell type, the transduced cells can be subjected to enrichment or selection assays specific to that cell type.
Enrichment and Amplification: After transduction and selection, the transduced cells are typically subjected to one or more rounds of enrichment or amplification to increase the representation of desirable capsids within the population. For the ATHENA I, researchers can use FACS to enrich EGPF positive cells.
Sequencing and Analysis: The enriched or amplified capsids are then characterized by sequencing their capsid genes. This can be done using next-generation sequencing (NGS) techniques to identify the specific capsid variants present in the selected population. Bioinformatic analysis is performed to determine the diversity, frequency, and characteristics of the identified capsids.
Functional Validation: Capsids with desired properties are further evaluated and validated for their functionality and performance in relevant assays or models. This can include assessing factors such as transduction efficiency, cell/tissue specificity, immune response, safety, and other relevant parameters.
Iterative Evolution: Based on the results obtained, the workflow can be iterated to generate new libraries with additional modifications or improvements. This allows for the progressive refinement and optimization of AAV capsids for specific applications.
Throughout the workflow, rigorous quality control measures will be implemented to ensure the integrity, purity, and safety of the AAV vectors and to minimize the risk of contamination or undesired characteristics.
It’s important to note that the specific details of the workflow may vary depending on the goals of the study, the available resources, and the specific techniques and assays employed.
