Department of Biochemistry, Chungnam National University, 99 Daehak-ro, Yuseong-gu, Daejeon 34134, Republic of Korea.
Molecules. 2023 Nov 17;28(22):7644. doi: 10.3390/molecules28227644.
Poly(amidoamine) (PAMAM) dendrimers have attracted considerable attention in the field of gene therapy due to their flexibility in introducing different functional moieties and reduced toxicity at low generations. However, their transfection efficiency remains a limitation. Therefore, an essential approach for improving their transfection efficiency as gene carriers involves modifying the structure of PAMAM by conjugating functional groups around their surface. In this study, we successfully conjugated an RRHRH oligopeptide to the surface of PAMAM generation 2 (PAMAM G2) to create RRHRH-PAMAM G2. This construction aims to condense plasmid DNA (pDNA) and facilitate its penetration into cell membranes, leading to its promising potential for gene therapy. RRHRH-PAMAM G2/pDNA complexes were smaller than 100 nm and positively charged. Nano-polyplexes can enter the cell and show a high transfection efficiency after 24 h of transfection. The RRHRH-PAMAM G2 was non-toxic to HeLa, NIH3T3, A549, and MDA-MB-231 cell lines. These results strongly suggest that RRHRH-PAMAM G2 holds promise as a gene carrier for gene therapy owing to its biocompatibility and ability to deliver genes to the cell.
聚（酰胺-胺）（PAMAM）树枝状聚合物由于其在引入不同功能部分方面的灵活性和在低代时毒性降低，在基因治疗领域引起了相当大的关注。然而，它们的转染效率仍然是一个限制。因此，作为基因载体，提高其转染效率的一个重要方法是通过在其表面连接官能团来修饰 PAMAM 的结构。在本研究中，我们成功地将 RRHRH 寡肽连接到 PAMAM 第二代（PAMAM G2）的表面，从而构建了 RRHRH-PAMAM G2。这种构建旨在浓缩质粒 DNA（pDNA）并促进其穿透细胞膜，因此具有作为基因治疗的潜在前景。RRHRH-PAMAM G2/pDNA 复合物的大小小于 100nm 且带正电荷。纳米多聚物可以进入细胞，并在转染 24 小时后显示出高转染效率。RRHRH-PAMAM G2 对 HeLa、NIH3T3、A549 和 MDA-MB-231 细胞系没有毒性。这些结果强烈表明，RRHRH-PAMAM G2 具有作为基因治疗基因载体的潜力，因为它具有生物相容性和将基因递送到细胞的能力。
