For example, based on experience, the higher GC content in cDNA of AAV, relatively higher titer but lower expression persistence in vivo.
Not my personal experience, but this paper is useful:
Faust, S. M., et al. (2013). "CpG-depleted adeno-associated virus vectors evade immune detection." J Clin Invest.
Due to their efficient transduction potential, adeno-associated virus (AAV) vectors are leading candidates for gene therapy in skeletal muscle diseases. However, immune responses toward the vector or transgene product have been observed in preclinical and clinical studies. TLR9 has been implicated in promoting AAV-directed immune responses, but vectors have not been developed to circumvent this barrier. To assess the requirement of TLR9 in promoting immunity toward AAV-associated antigens following skeletal muscle gene transfer in mice, we compared immunological responses in WT and Tlr9-deficient mice that received an AAV vector with an immunogenic capsid, AAVrh32.33. In Tlr9-deficient mice, IFN-gamma T cell responses toward capsid and transgene antigen were suppressed, resulting in minimal cellular infiltrate and stable transgene expression in target muscles. These findings suggest that AAV-directed immune responses may be circumvented by depleting the ligand for TLR9 (CpG sequences) from the vector genome. Indeed, we found that CpG-depleted AAVrh32.33 vectors could establish persistent transgene expression, evade immunity, and minimize infiltration of effector cells. Thus, CpG-depleted AAV vectors could improve outcome of clinical trials of gene therapy for skeletal muscle disease.
Thanks Dr Waddington,
Except delete CpG island in cDNA, is ~65% of CG% highly possible to enhance AAV titer, however ＞65% of CG% is also highly possible to induce TLR9 immune response. What the percentage of CG content in cDNA do you recommend?
I am using AAV9 and design its codon-optimized cDNA.
I am also worring about the patent issue of AAV9 and its purification effect if I used affinity chromotography.
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