A novel and highly efficient AAV6 mutant

作者：Wang, Feifei; Huang, Can; Cao, Jinjing; Liu, Xiaomei^*; Wang, Dongxin; Zhang, Chun; Chen, Qin

来源：Virus Genes, 2018, 54(2): 165-171.

DOI：10.1007/s11262-017-1531-2

• AAV6
• Fusion PCR
• Site mutation
• Mutant
• Transduction efficiency
• Gene therapy

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.

• 出版日期2018-4
• 单位中国科学院; 中国科学院苏州生物医学工程技术研究所; 上海大学; 中国科学技术大学