The Role of Proteoglycans in Retroviral Mediated Gene Transfer

9800617 Yarmush这项SGER提案涉及提高通常使用重组逆转录病毒转移到哺乳动物细胞的有效性。在之前的工作中，PI已经发现由病毒产生细胞产生的糖蛋白抑制逆转录病毒介导的基因转移。降解这些蛋白多糖的酶提高了将基因转移到哺乳动物细胞的生产率。由于蛋白多糖和病毒颗粒的大小相似，物理分离似乎不太可能。这项提议的总体目标是更好地了解蛋白多糖在抑制过程中的作用。这将通过以下方式完成：分离和纯化蛋白多糖，研究它们的影响，以及它们的蛋白质和多糖片段对转移过程的影响，测量这些抑制物与靶细胞和逆转录病毒的结合。确定抑制物片段和负责抑制作用的细胞受体。建立抑制过程的模型更好地了解抑制机制及其范围应有助于发现解决问题的途径。已经考虑过的一条这样的途径是加入破坏蛋白多糖的基因。***

9800617 Yarmush This SGER proposal deals with increasing the effectiveness often transfer to mammalian cells using recombinant retroviruses. In previous efforts, the PIs have found that glycoproteins produced by virus producer cells inhibit retroviral-mediated gene transfer. Enzymes that degrade these proteoglycans enhance the productivity of the gene transfer to mammalian cells. Since the size of the proteoglycans and the viral particles are similar, physicals separations seem unlikely. The overall goal of this proposal is to better understand the role of the proteoglycans in the inhibition process. This will be accomplished by: Isolating and purifying the proteoglycans and studying their impact, and the impact of their protein and glycan fragments on the transfer process Measuring the binding of these inhibitors to both the target cells and the retrovirus. Identify the inhibitor fragment and the cell receptor responsible for the inhibition. Develop a model of the inhibition process A better understanding of the inhibition mechanism and its scope should facilitate discovery of routes to remedy the problem. One such route already considered involves incorporation of genes for proteoglycan destruction. ***