AAV Production and library
ATHENA AAV Capsid Screening Platform
ATHENA: AAVnerGene's Tissue-specific, Highly-transductive and Expressive New AAVs.
ATHENA is a two-step AAV selection Platform. First step, use our ATHENA I platform to select the best well known AAV vectors for each cell type. Second step, use our high complexity, random peptide ATHENA II AAV library to further optimize the selected capsids in ATHENA I.
ATHENA II AAV Capsid Library
Libraries displaying random peptides on the surface of AAV are powerful tools for screening of target-specific gene therapy vectors. Random peptide sequences have been inserted into defined sites of the viral capsid, such as in the heparin binding domain of the AAV2 capsid (at capsid residue R588), via ligation of degenerate oligonucleotides into the cap ORF.
Recently studies have highlighted several loop regions on the surface of AAV capsid influence the transduction properties of AAV vectors. Engineer of these surface loops would allow us to develop novel AAV capsid variants.
AAVnerGene's ATHENA II AAV capsid libraries are developed based on random peptides display technology. By analyzing the sequences of different AAV capsids, we found many sites which are variable and can be replaced or inserted with the random peptides. Some of those sites can be used to introduce random peptides without significantly affecting AAV packaging.
Random peptide insert sites
Generation of AAV libraries
AAV vectors are built on the selected AAV capsids and selected IRs. Two BaeI sites are introduced at IRs and the Random peptides are inserted into the BaeI sites. AAVnerGene developed a new technology that can efficiently introduce the random peptides into selected sites. We also developed a new process that significanlty increase the library complexity. With these novel technologies, we are able to generate a complexity of 1e8 AAV capsid library in one ligation process. We can easily construct a library with complexity at 1E9 that will greatly enhance the chance to select the best AAV serotype for a specific cell type.
Complexity of ATHENA II AAV capsid library
With our novel library construction strategies, we can easily construct a library with complexity at 1E9. ATHENA II also can group all the sub-libraries together and make a AAV capsid library with complexity over 1E11. With the high complexity, ATHENA II will greatly enhance the chance to select the best AAV serotype for a specific cell type
Custom AAV Capsid Libraries
For the custom AAV capsid libraries, customers can choose their library templates, length of random peptides, insert sites and library complexity. Please contact us for more information.
Procedures of making custom AAV capsid selection kits
Price and Time
Construction of backbone plasmids: 1-2 week.
Construction of random peptide library: 1-2 week
Production of AAV library: 1-2 weeks.
Quality control: 2-3 weeks
Total time: 6~9 week.