|作者：||Li Hua, Zhang Feng-Lan, Shi Wen-Jie, Bai Xue-Jia, Jia Shu-Qin, Zhang Chen-Guang, Ding Wei|
1Department of Oral and Maxillofacial-Head and Neck Oncology, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.
2National Institutes for Food and Drug Controls, Beijing, China.
3Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China; Department of Medical Genetics, Capital Medical University, Beijing, China.
4Department of Medical Genetics, Capital Medical University, Beijing, China.
5Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
6Department of Medical Genetics, Capital Medical University, Beijing, China; Beijing Institute of Brain Disorders, Beijing, China.
|刊名：||PloS one, 2015, Vol.10 (6), pp.e0129013|
|原始语种摘要：||The technology of virus-based genetic modification in tissue engineering has provided the opportunity to produce more flexible and versatile biomaterials for transplantation. Localizing the transgene expression with increased efficiency is critical for tissue engineering as well as a challenge for virus-based gene delivery. In this study, we tagged the VP2 protein of type 2 adeno-associated virus (AAV) with a 3×FLAG plasmid at the N-terminus and packaged a FLAG-tagged recombinant AAV2 chimeric mutant. The mutant AAVs were immobilized onto the tissue engineering scaffolds with crosslinked anti-FLAG antibodies by N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP). Cultured cells were seeded to scaffolds to form 3D transplants, and then tested for viral transduction both in vitro and in... vivo. The results showed that our FLAG-tagged AAV2 exerted similar transduction efficiency compared with the wild type AAV2 when infected cultured cells. Following immobilization onto the scaffolds of PLGA or gelatin sponge with anti-FLAG antibodies, the viral mediated transgene expression was significantly improved and more localized. Our data demonstrated that the mutation of AAV capsid targeted for antibody-based immobilization could be a practical approach for more efficient and precise transgene delivery. It was also suggested that the immobilization of AAV might have attractive potentials in applications of tissue engineering involving the targeted gene manipulation in 3D tissue cultures.|