Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
Biomaterials. 2010 May;31(14):4194-203. doi: 10.1016/j.biomaterials.2010.01.116. Epub 2010 Feb 16.
Ternary nanoparticles with negatively charged surface were prepared by coating single-stranded oligonucleotides (5'-C(10)A(20)-3') on histidine-conjugated polyallylamine (PAA-HIS)/DNA complexes for gene delivery. Characterization of PAA-HIS/DNA/oligonucleotide complexes demonstrated that nanoparticles possessed the negative surface charge -27 mV and size of around 100 nm when the molar ratio of oligonucleotide/PAA-HIS exceeded 1.5. The negatively charged oligonucleotide-coated PAA-HIS/DNA complexes could be entirely internalized by the living HeLa cells to exhibit high gene expression with low cytotoxicity and the resistance against erythrocyte agglutination and serum inhibition. Since the gene expression of PAA-HIS/DNA complexes was significantly inhibited by coating other polyanions and oligonucleotides, the ternary PAA-HIS/DNA/deoxyadenosine-rich oligonucleotide complexes were uptaken by specific receptor-mediated process. Additionally, the deposition of a layer of oligonucleotides onto the binary PAA-HIS/DNA complexes could effectively transfect various types of cells including HEK-293, HepG2 and Hs68 cells, indicating the technology of coating specific oligonucleotides on PAA-HIS/DNA complexes or other cationic binary DNA complexes might facilitate the use of nanoparticles for safe and efficient gene delivery and eventual therapy.
采用带负电荷的单链寡核苷酸（5'-C（10）A（20）-3'）对组氨酸缀合的聚烯丙胺（PAA-HIS）/DNA 复合物进行包覆，制备出具有负表面电荷的三元纳米粒子，用于基因传递。当寡核苷酸/PAA-HIS 的摩尔比超过 1.5 时，PAA-HIS/DNA/寡核苷酸复合物的特性表明，纳米粒子具有负表面电荷-27 mV 和约 100 nm 的粒径。带负电荷的寡核苷酸包覆的 PAA-HIS/DNA 复合物可被活的 HeLa 细胞完全内化，表现出低细胞毒性和抗红细胞聚集及血清抑制的高基因表达。由于 PAA-HIS/DNA 复合物的基因表达被其他聚阴离子和寡核苷酸的包覆显著抑制，因此三元 PAA-HIS/DNA/去氧腺苷富寡核苷酸复合物通过特定的受体介导过程被摄取。此外，在二元 PAA-HIS/DNA 复合物上沉积一层寡核苷酸可以有效地转染包括 HEK-293、HepG2 和 Hs68 在内的各种类型的细胞，表明在 PAA-HIS/DNA 复合物或其他阳离子二元 DNA 复合物上包覆特定寡核苷酸的技术可能有助于安全有效地进行基因传递和最终治疗。
