Environmental Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141; Department of Biosystems and Bioengineering, KRIBB School of Biotechnology, University of Science and Technology (UST), Daejeon 34113, Korea.
Environmental Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Korea.
BMB Rep. 2020 Aug;53(8):442-447. doi: 10.5483/BMBRep.2020.53.8.024.
The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model. [BMB Reports 2020; 53(8): 442-447].
将基因非病毒递送至巨噬细胞（即难转染细胞）是一个挑战。在这项研究中，CpG 无质粒和小质粒（pCGfd-GFP）的电穿孔显示出在 Raw 264.7 和原代骨髓来源的巨噬细胞转染中具有高转染效率、可持续的转基因表达和良好的细胞活力。使用编码抗 EGFR 单链 Fv 与 Fc 融合的 pCGfd 载体的非病毒方法生成了分泌抗 EGFR scFv-Fc 的巨噬细胞。这些巨噬细胞通过抗体依赖性机制有效地吞噬表达 EGFR 的肿瘤细胞，这通过使用 EGFR 敲除肿瘤细胞的实验得到了证明。最后，在异种移植小鼠模型中，经皮注射分泌抗 EGFR scFv-Fc 的巨噬细胞抑制了肿瘤生长。[BMB 报告 2020；53(8): 442-447]。
