Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory for Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Aptamer Engineering Center of Hunan Province, Hunan University , Changsha, Hunan 410082, China.
Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, University Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida , Gainesville, Florida 32611-7200, United States.
J Am Chem Soc. 2018 Jan 10;140(1):2-5. doi: 10.1021/jacs.7b08518. Epub 2017 Dec 27.
Enhanced targeted gene transduction by AAV2 vectors is achieved by linking the vector to multiple sgc8 aptamers, which are selective for cell membrane protein PTK7. Aptamer molecules are conjugated to multiple sites on a DNA dendrimer (G-sgc8), which is then linked to AAV2 via a dithiobis(succinimidyl propionate) cross-linker containing a disulfide group, which can facilitate the release of AAV2 vectors by reaction with the reduced form of intracellular glutathione. The G-sgc8-AAV2 vectors showed a 21-fold enhancement in binding affinity and an enhanced ability to protect sgc8 aptamers against nuclease degradation to cells expressing PTK7 compared to single aptamer-AAV2 conjugates. The transduction efficiency was tested by loading AAV2 with the gene for green fluorescent protein. Therefore, this modified recombinant vector is an attractive and promising tool for targeted biomedical applications.
通过将载体与多个针对细胞膜蛋白 PTK7 的 sgc8 适体连接，实现了 AAV2 载体的增强靶向基因转导。适体分子连接到 DNA 树枝状大分子（G-sgc8）的多个位点上，然后通过含有二硫键的二硫代琥珀酰亚胺基丙酸（Dithiobis(succinimidyl propionate)）交联剂与 AAV2 连接，该二硫键可以通过与细胞内还原型谷胱甘肽反应促进 AAV2 载体的释放。与单个适体-AAV2 缀合物相比，G-sgc8-AAV2 载体与表达 PTK7 的细胞的结合亲和力提高了 21 倍，并且能够增强保护 sgc8 适体免受核酸酶降解的能力。通过装载携带绿色荧光蛋白基因的 AAV2 来测试转导效率。因此，这种改良的重组载体是一种有吸引力和有前途的靶向生物医学应用工具。
