Virus Genes

, Volume 54, Issue 2, pp 165–171 | Cite as

A novel and highly efficient AAV6 mutant

  • Feifei Wang
  • Can Huang
  • Jinjing Cao
  • Xiaomei Liu
  • Dongxin Wang
  • Chun Zhang
  • Qin Chen
Article
  • 123 Downloads

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.

Keywords

AAV6 Fusion PCR Site mutation Mutant Transduction efficiency Gene therapy 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 311101101).

Author contributions

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable guidelines by the Institute’s Animal Ethics Committee No.F.1-53/2012-13-J.D. (Res) for the care and use of animals were followed.

Informed consent

All authors read and approved the manuscript.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  • Feifei Wang
    • 1
    • 2
  • Can Huang
    • 1
    • 2
  • Jinjing Cao
    • 2
    • 3
  • Xiaomei Liu
    • 2
  • Dongxin Wang
    • 2
    • 3
  • Chun Zhang
    • 2
  • Qin Chen
    • 1
  1. 1.Shanghai UniversityShanghaiChina
  2. 2.Suzhou Institute of Biomedical Engineering and TechnologyChinese Academy of SciencesSuzhouChina
  3. 3.University of Science and Technology of ChinaHefeiChina

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