AAV Capsid Library Construction

AAV capsid libraries are collections of diverse AAV variants with engineered or randomized capsid sequences. These libraries are used to screen for novel AAV capsids with specific properties, such as improved transduction efficiency, tissue targeting, or immune evasion. 

The design and construction of an AAV capsid library are crucial for selecting the most suitable AAV capsid for a specific application. The library should be diverse and complex enough to ensure that all possible capsid variants are represented and that the screening process can identify the most efficient and specific capsids.

AAVnerGene offers two different AAV capsid libraries and its related services:

  • AAV Capsid Library(I)is build on our ATHENA I platform. Itis a known capsid library used to systematically compare and select the best AAV serotypes or variants. In the ATHENA I library, each capsid variant corresponds to three different DNA barcoded genomes with the same reporter. The unique barcode is located between the coding sequence and the Poly-A signal. During AAV production, one capsid and assigned 3 barcode plasmids were co-transfected to make one specific capsid AAV vector. All the AAV vectors were harvested, purified, and tittered, either pool or separately. By comparing the DNA or RNA levels of the reporter gene with NGS targeting the BCs, researchers can determine which AAV capsid variant is the most efficient for their specific application.
ATHENA I Known Capsid Library
  • AAV Capsid Library(II) is build on our ATHENA II platform. It is a random peptide insert library used to evolute novel AAV capsid with tissue specific tropism.  In this library, random peptide sequences are inserted into variable sites of the AAV capsid protein. These peptides can potentially alter the properties of the capsid, such as its tropism, transduction efficiency, and immunogenicity, and thus affect its utility for gene therapy applications. By screening the library, researchers can identify novel capsids with improved properties and select the ones that are optimal for their specific therapeutic applications. 
AAV Capsid Library(II)
  AAV Capsid Library(I) AAV Capsid Library(II)
Library type ATHENA I
Known AAV capsid collection library
ATHENA II
Random peptide insert library
Purpose Evaluate different AAV capsids from known serotypes/variants Create and evolve novel AAV capsids
Complexity Low(~1000 different AAV variants) High(>1e9 variants)
Reporter  CAG-EGFP CAG/P40-Cap
DNA barcodes One capsid corresponding 3 different specific DNA barcodes Random peptides themselves
Production condition Triple-plasmid transfection as usual Low copy number of library plasmid plus normal pHelper and pRep plasmid
Selection method NGS (~100M)  NGS(usually>100M)
Premade library Yes Yes
Custom library services Yes Yes
Selection services Yes Yes
 Prices  Based on request  Based on request