Abstract
Adeno-associated virus has been gaining prominence in its use as a highly secure virus gene vector with low immunogenicity in the field of human gene therapy. However, wild-type adeno-associated virus sometimes has low transduction efficiency for certain tissues or cells both in vivo and in vitro. Thus, achieving the desired level of expression often requires a large dose. Large doses of viral injection in clinical applications will not only trigger the body’s immune response but will come at a high production cost. To improve the transduction efficiency of adeno-associated virus 6 (AAV6), we herein used fusion PCR to mutate a specific amino acid of the VP2 region of the wild-type AAV6 (AAV6-WT) and obtained AAV6-S663L, AAV6Y705 + 731F + T492A, AAV6Y705 + 731F + T492 V + S663 V and so on. We concluded that AAV6-S663L was the most efficient AAV6 mutant. When HEK293 cells were infected in vitro with a virus at a multiplicity of infection value of 1000, the transduction rate of AAV6-WT was only 43.8%, while that of AAV6-S663L was 83.9%. This highly efficient AAV6 mutant is highly significant for the future use of AAV6 in gene therapy.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant No. 311101101).
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Conceived and designed the experiments: FFW, CH. Performed the experiments: FFW, CH, JJC. Analysed the data: FFW, XML, DXW. Wrote the paper: FFW, CZ, QC. Read and approved the manuscript: FFW, CH, JJC, DXW, XML, CZ, QC.
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Wang, F., Huang, C., Cao, J. et al. A novel and highly efficient AAV6 mutant. Virus Genes 54, 165–171 (2018). https://doi.org/10.1007/s11262-017-1531-2
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DOI: https://doi.org/10.1007/s11262-017-1531-2