Earticle

초록

영어

Background: The ability of adeno-associated viruses (AAVs) to transduce various cell types with minimal immune responses renders them prominent vectors for gene editing (GE), with different AAV serotypes exhibiting distinct transduction efficiencies due to their specific cellular tropism. However, detailed molecular processes of AAV infection and penetration, as well as the optimal serotype for specific purposes, remain poorly understood. Porcine models are widely used in research benefitting both human and livestock due to anatomical and physiological similarities to humans. Methods: Transduction efficiencies of 18 AAV serotypes (AAV1–9, 6.2, rh10, DJ, DJ/8, PHP.eB, PHP.S, 2-retro, 2-QuadYF, and 2.7m8) were evaluated in immortalized porcine lung epithelial cells (pLCsImt) and pulmonary alveolar macrophages 3D4/31 (PAMs 3D4/31). Results: We found AAV2, DJ, and 2.7m8 to be the most effective in both cell types. The highest enhanced green fluorescent protein expression of 52.46 ± 2.4% in pLCsImt and 64.08 ± 2.4% in PAMs 3D4/31 was observed for AAV2, while negligible transduction was observed for AAV4, rh10, DJ, PHP.eB, PHP.S, and 2-retro. AAV-DJ showed superior transduction efficiency in PK-15, as compared to AAV2 and 2.7m8. Results emphasize the cell type-specific nature of AAV serotype transduction efficiencies. Notably, AAV2 was most effective in both lung and macrophage cells, whereas AAV-DJ was more effective in renal cells. Conclusions: Our findings suggest that AAV2 was identified as the most efficient serotype for transducing pLCsImt and PAMs 3D4/31, compare to the PK-15 cells. Understanding cell type-specific preferences of AAV serotypes offer crucial insight for tailoring AAV vectors to specific tissue and optimizing genome editing strategies, with potential implications for the advancement of personalized medicine and development of treatments for human and livestock.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Cell culture
Immortalization of pLCs using lentivirus particles
Single cell sorting by FACS
AAVs and in vitro transduction
Transduction efficiency
Real-time qPCR
Statistical analysis
RESULTS
AAV serotypes show different transduction efficiencies in immortalized porcine lung epithelial cells
AAV serotypes show comparable transduction efficiency in PAMs 3D4/31 to pLCsImt
Transduction efficiency of individual AAV serotypes in porcine-derived cell types
DISCUSSION
CONCLUSION
SUPPLEMENTARY MATERIALS
REFERENCES

키워드

저자

• Won Seok Ju [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Seokho Kim [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ] Corresponding Author
• Areum Choi [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Jae-Yeong Lee [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Haesun Lee [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Jingu No [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Seunghoon Lee [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Keonbong Oh [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]
• Jae Gyu Yoo [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Korea ]

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2026

• 십진분류

  KDC 527 DDC 636