Skoumal Michael, Seidlits Stephanie, Shin Seungjin, Shea Lonnie
Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan.
Department of Bioengineering, University of California, Los Angeles, Los Angeles, California.
Biotechnol Bioeng. 2016 Sep;113(9):2033-40. doi: 10.1002/bit.25961. Epub 2016 Mar 6.
Gene delivery from biomaterial scaffolds has been employed to induce the expression of tissue inductive factors for applications in regenerative medicine. The delivery of viral vectors has been described as reflecting a balance between vector retention and release. Herein, we investigated the design of hydrogels in order to retain the vector at the material in order to enhance transgene expression. Poly(ethylene-glycol) (PEG) hydrogels were modified with poly-l-lysine (PLL) to non-covalently bind lentivirus. For cells cultured on the hydrogels, increasing the PLL molecular weight from 1 to 70 kDa led to increased transgene expression. The incubation time of the virus with the hydrogel and the PLL concentration modulated the extent of virus adsorption, and adsorbed virus had a 20% increase in the half-life at 37°C. Alternatives to high molecular weight PLL were identified through phage display technology, with peptide sequences specific for the VSV-G ectodomain, an envelope protein pseudotyped on the virus. These affinity peptides could easily be incorporated into the hydrogel, and expression was increased 20-fold relative to control peptide, and comparable to levels observed with the high molecular weight PLL. The modification of hydrogels with affinity proteins or peptides to bind lentivirus can be a powerful strategy to enhance and localized transgene expression. Biotechnol. Bioeng. 2016;113: 2033-2040. © 2016 Wiley Periodicals, Inc.
生物材料支架的基因递送已被用于诱导组织诱导因子的表达，以应用于再生医学。病毒载体的递送被描述为反映了载体保留和释放之间的平衡。在此，我们研究了水凝胶的设计，以便将载体保留在材料上，从而增强转基因表达。聚（乙二醇）（PEG）水凝胶用聚-L-赖氨酸（PLL）进行修饰，以非共价结合慢病毒。对于在水凝胶上培养的细胞，将PLL分子量从1 kDa增加到70 kDa会导致转基因表达增加。病毒与水凝胶的孵育时间和PLL浓度调节了病毒吸附程度，并且吸附的病毒在37°C下的半衰期增加了20%。通过噬菌体展示技术鉴定了高分子量PLL的替代物，其具有针对VSV-G胞外域的肽序列，VSV-G是一种在病毒上假型化的包膜蛋白。这些亲和肽可以很容易地掺入水凝胶中，并且表达相对于对照肽增加了20倍，与高分子量PLL观察到的水平相当。用水凝胶与亲和蛋白或肽修饰以结合慢病毒可以是增强和定位转基因表达的有力策略。生物技术与生物工程。2016年；113：2033 - 2040。©2016威利期刊公司。
