AAV2 Reference-4.3kb

The AAV2 reference-4.3kb material contains a 4.3kb AAV-CMV-SV40I-EGFP-WPRE-hGHpolyA genome (with a 1.5kb LacZ) in AAV2 capsid, with a genome titer (EGFP targeting) of 1X10^13 VG/ml. This reference was produced using the AAVone system with the pAAVone-AAV2-CMV-EGFP-4.3 kb plasmid, which was generated by inserting a 1.5 kb LacZ sequence as a stuffer upstream of the CMV promoter in the pAAVone-AAV2-CMV-EGFP-2.8 kb plasmid. AAV2 is one of the most well-studied and widely used serotypes of adeno-associated virus. It has been a workhorse for gene therapy research due to its well-characterized biology and its ability to transduce a broad range of tissues, including the central nervous system (CNS), liver, and retina. AAV2 was the first AAV serotype to be cloned and is often used as a reference or baseline for comparing other AAV serotypes.

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Background

AAV references are crucial for qPCR-based vector genome quantification and fundamental for ensuring accuracy in viral vector research and gene therapy development. Our AAV reference materials undergo extensive characterization, including full ratio (mass photometry), genome titer (ddPCR), capsid titer (ELISA), purity (SDS-PAGE silver staining), and tests for bioburden, endotoxin, and mycoplasma. Quantification is based on reference material from ATCC. These thoroughly analyzed particles provide precise genome titers, well-defined full-to-empty capsid ratios, and accurate capsid titers. This comprehensive characterization is essential for standardizing measurements, ensuring consistency across experiments, and delivering reliable data for gene therapy applications and other research involving AAV vectors.
AAV references (Truly-Empty) are produced by transfection of a single mini-pHelper-AAV plasmid with the corresponding serotypes. AAV references (Full) and AAV references (Byproduct-Empty) are generated using the AAVone system, with the corresponding pAAVone plasmids carrying AAV-CMV-SV40I-EGFP-WRRE-hGHpolyA genomes.