Transduction of Mouse Retina By Insect Cell Packaged Recombinant adeno-associated Viruses and Their Mutants Via Intravitreal Injection

Abstract

Aim: Adeno-associated virus (AAV) is the most preferred gene therapy vector. The purpose of our research is to compare the infection tropism and gene expression efficiency of vitreous injection of recombinant AAVs (rAAVs) and their capsid mutants in mouse retina. Materials & methods: We packaged wild-type rAAV2/2,6,8,9 and their capsid mutants carrying EGFP expression cassette using insect cells. The gene expression profiles of rAAVs and their mutants in mouse retina were evaluated by optical imaging of retinal tissue flat mount and cryosections. Results & conclusion: The results showed that rAAV2 and rAAV2-Y444F mainly targeted retinal ganglion cell; rAAV8, rAAV8-Y733F, rAAV9 and mutants had obvious EGFP expression in retinal pigment epithelium cells. Compared with the wild-type rAAVs, capsid mutants have an improved transduction efficiency in mouse retina cells.

Keywords:

• adeno-associated viruses
• intravitreal injection
• mouse retina
• transduction efficiency
• tropism

Author contributions

C Zhang and T Li designed the experiments, Z Wei and X Liu conducted the experiments, Z Wei and X Liu analyzed the experimental data. The first draft of the manuscript was written by X Li, Q Zhou and J Wu. J Wu provides financial support for research. All authors read and approved the final manuscript.

Acknowledgments

The authors thanked D Wang for technical support. The authors thanked X Liu and Z Wei for their reading of the manuscript and helpful suggestions.

Financial & competing interests disclosure

This work was supported by grants from the Suzhou Municipal Science and Technology Program (Grant No. SS201761), and the Natural Science Foundation of Hunan Province (NO. 2020JJ4143). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The mice experimentation protocol was approved by the Animal Welfare and Ethics Committee of Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences under permit number SIBET-2019-A-06.