Effects of trehalose polycation end-group functionalization on plasmid DNA uptake and transfection.
Effects of trehalose polycation end-group functionalization on plasmid DNA uptake and transfection.
复制标题
海藻糖多阳离子终端官能化对质粒DNA摄取和转染的影响。
DOI：
10.1021/bm300471n
复制
发表时间：
2012-08-13
影响因子：
6.2
通讯作者：
Reineke, Theresa M.
中科院分区：
文献类型：
--
作者：
Anderson, Kevin;Sizovs, Antons;Cortez, Mallory;Waldron, Chris;Haddleton, D. M.;Reineke, Theresa M.
在这项研究中，我们已经合成了六个类似物的海藻糖-五亚乙基六胺glycopolymer（Tr 4），含有（1A）金刚烷，（1B）羧基，（1C）炔基-低聚乙烯胺，（1D）叠氮基海藻糖，（1 E）辛基，或（1F）低聚乙烯胺端基，并评估了聚合物端基化学对这些系统的能力的影响，以结合，压缩，并提供pDNA在培养的HeLa细胞。在一锅叠氮-炔环加成反应中合成聚合物，并采用Carothers方程进行逐步增长聚合，以产生一系列具有相似聚合度的聚合物。在聚合结束时加入过量的封端单体以使官能化效率最大化，这用GPC、NMR和MALDI-TOF进行评价。聚合物都被发现以类似的低N/P比结合和压缩pDNA，并与质粒DNA形成聚合复合物。在血清存在下完成的复合物内化和转染测定中，不同端基结构的影响最明显，分别通过流式细胞术和荧光素酶基因表达测定。用羧基（1B）（N/P = 7）、辛炔（1 E）（N/P = 7）和低聚亚乙基胺（1F）（N/P = 7）封端的Tr 4聚合物作为多聚物进入细胞，并表现出最高水平的荧光，这是由标记的报告质粒产生的。类似地，用羧基（1 E，N/P = 7）、辛基（1 E，N/P = 15）和特别是低聚亚乙基胺基团（F，N/P = 15）末端官能化的聚合物在血清存在下产生显著更高的报告基因表达。该研究深入了解了聚合物化学中非常细微的结构变化（例如端基）如何在用于pDNA递送的聚合物的生物递送效率和转基因表达中产生非常显著的差异。
In this study, we have synthesized six analogs of a trehalose-pentaethylenehexamine glycopolymer (Tr4) that contain (1A) adamantane, (1B) carboxy, (1C) alkynyl-oligoethyleneamine, (1D) azido trehalose, (1E) octyl, or (1F) oligoethyleneamine end groups and evaluated the effects of polymer end group chemistry on the ability of these systems to bind, compact, and deliver pDNA in cultured HeLa cells. The polymers were synthesized in one-pot azide-alkyne cycloaddition reactions with an adaptation of the Carothers equation for step-growth polymerization to produce a series of polymers with similar degrees of polymerization. An excess of end-capping monomer was added at the end of the polymerizations to maximize functionalization efficiency, which was evaluated with GPC, NMR and MALDI-TOF. The polymers were all found to bind and compact pDNA at similarly low N/P ratios and form polyplexes with plasmid DNA. The effects of the different end group structures were most evident in the polyplex internalization and transfection assays completed in the presence of serum, as determined by flow cytometry and luciferase gene expression respectively. The Tr4 polymers end-capped with carboxyl groups (1B) (N/P = 7), octyne (1E) (N/P = 7), and oligoethyleneamine (1F) (N/P = 7), were taken into cells as polyplex and exhibited the highest levels of fluorescence, resulting from labeled reporter plasmid. Similarly, the polymers end-functionalized with the carboxyl groups (1E at N/P = 7), octyl groups (1E at N/P = 15) and, in particular, the oligoethyleneamine groups (F at N/P = 15) yielded dramatically higher reporter gene expression in the presence of serum. This study yields insight into how very subtle structural changes in the polymer chemistry such as end groups can yield very significant differences in the biological delivery efficiency and transgene expression of polymers used for pDNA delivery.
登录
查看更多内容
影响因子：
13.5
作者：
Jung, CR;Yoo, J;Iml, DS
通讯作者：
Iml, DS
DNA/核蛋白/聚阳离子与基因传递三联体的相互作用
影响因子：
3.5
作者：
Shen, Yuan;Peng, Hui;Wu, Chuanbin
通讯作者：
Wu, Chuanbin
影响因子：
14
作者：
Srinivasachari, Sathya;Liu, Yemin;Reineke, Theresa M.
通讯作者：
Reineke, Theresa M.
影响因子：
14
作者：
Bhise, Nupura S.;Gray, Ryan S.;Sunshine, Joel C.;Htet, Soe;Ewald, Andrew J.;Green, Jordan J.
通讯作者：
Green, Jordan J.
影响因子：
10.8
作者：
Park, MR;Han, KO;Cho, CS
通讯作者：
Cho, CS