Sun Liping, Xiong Yunjing, Bashan Anat, Zimmerman Ella, Shulman Daube Shirley, Peleg Yoav, Albeck Shira, Unger Tamar, Yonath Hagith, Krupkin Miri, Matzov Donna, Yonath Ada
Department of Biomaterials, College of Materials, Xiamen University, 422, Siming South Road, Xiamen 361005 (China).
Weizmann Institute of Science, 234 Herzl Street, Rehovot 7610001 (Israel).
Chembiochem. 2015 Jul 6;16(10):1415-9. doi: 10.1002/cbic.201500205. Epub 2015 May 26.
We have developed a collagen-mRNA platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures. A collagen mimic was constructed according to a recombinant method and was used as scaffold for translating mRNA chains into proteins. Cysteines were genetically inserted into the collagen chain at positions allowing efficient ribosome translation activity while minimizing mRNA misfolding and degradation. Enhanced green fluorescence protein (eGFP) mRNA bound to collagen was successfully translated by cell-free Escherichia coli ribosomes. This system enabled an accurate control of specific protein synthesis by monitoring expression time and level. Luciferase-mRNA was also translated on collagen scaffold by eukaryotic cell extracts. Thus we have demonstrated the feasibility of controllable protein synthesis on collagen scaffolds by ribosomal machinery.
我们开发了一种用于可控蛋白质生产的胶原蛋白 - mRNA平台,旨在减少与常用mRNA疗法以及胶原蛋白皮肤愈合程序相关的问题。根据重组方法构建了一种胶原蛋白模拟物,并将其用作将mRNA链翻译成蛋白质的支架。半胱氨酸被基因插入到胶原蛋白链中,这些位置既能保证高效的核糖体翻译活性,又能使mRNA的错误折叠和降解最小化。与胶原蛋白结合的增强型绿色荧光蛋白(eGFP)mRNA被无细胞的大肠杆菌核糖体成功翻译。该系统通过监测表达时间和水平实现了对特定蛋白质合成的精确控制。荧光素酶 - mRNA也通过真核细胞提取物在胶原蛋白支架上进行了翻译。因此,我们证明了通过核糖体机制在胶原蛋白支架上进行可控蛋白质合成的可行性。