Location of a single histidine within peptide carriers increases mRNA delivery.
Location of a single histidine within peptide carriers increases mRNA delivery.
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在肽载体内的单个组氨酸的位置增加mRNA递送。
DOI：
10.1002/jgm.3295
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Mixson AJ
中科院分区：
文献类型：
--
作者：
He J;Xu S;Leng Q;Mixson AJ
此前，我们确定四支链组氨酸-赖氨酸（HK）肽是质粒和小干扰RNA的有效载体。在本研究中，我们比较了几种分支 HK 载体，特别是两种密切相关的 H3K4b 和 H3K(+H)4b 肽作为 mRNA 载体的能力。 H3K(+H)4b 肽与其母体类似物 H3K4b 的不同之处在于每个分支中仅存在一个组氨酸。在固相肽合成仪上合成了一系列具有不同序列的四支链HK肽。研究了这些肽将表达荧光素酶的 mRNA 携带至 MDA-MB-231 细胞的能力。通过凝胶阻滞和肝素置换测定，检查了 HK 复合物的稳定性。我们通过流式细胞术和荧光显微镜测定了 HK 复合物的细胞内摄取。通过动态光散射测量了多种介质中聚合物复合物的尺寸和多分散指数。 H3K(+H)4b-mRNA 复合物转染的 MDA-MB-231 细胞表达的荧光素酶水平比 H3K4b 复合物高 10 倍。通过凝胶阻滞和肝素置换测定，H3K(+H)4b 复合物表现出比 H3K4b 更高的稳定性。与 H3K4b 相比，酸性内体中 H3K(+H)4b 复合物的细胞内摄取和共定位也显着增加。与 H3K(+H)4b 类似，一些在其分支的第二个结构域中具有额外组氨酸的 HK 类似物是 mRNA 的有效载体。当与 DOTAP 脂质体结合时，H3K(+H)4b 在 mRNA 递送方面具有协同作用。 H3K(+H)4b 是比 H3K4b 更有效的 mRNA 载体。机理研究表明 H3K(+H)4b 聚合复合物比 H3K4b 聚合复合物更稳定。 H3K(+H)4b 形成的脂多聚复合物显着增加了 mRNA 转染。在特定位置添加一个额外组氨酸的支链 HK mRNA 复合物显示出其细胞摄取、内体内积累以及转染效率的显着改善。那些缺乏形成 mRNA 多聚体的额外组氨酸的 HK 载体稳定性降低，导致转染率较低。
Previously, we determined that four‐branched histidine‐lysine (HK) peptides were effective carriers of plasmids and small interfering RNA. In the present study, we compared several branched HK carriers and, in particular, two closely‐related H3K4b and H3K(+H)4b peptides for their ability as carriers of mRNA. The H3K(+H)4b peptide differed from its parent analogue, H3K4b, by only a single histidine in each branch. A series of four‐branched HK peptides with varied sequences was synthesized on a solid‐phase peptide synthesizer. The ability of these peptides to carry mRNA expressing luciferase to MDA‐MB‐231 cells was investigated. With gel retardation and heparin displacement assays, the stability of HK polyplexes was examined. We determined the intracellular uptake of HK polyplexes by flow cytometry and fluorescence microscopy. The size and polydispersity index of the polyplexes in several media were measured by dynamic light scattering. MDA‐MB‐231 cells transfected by H3K(+H)4b‐mRNA polyplexes expressed 10‐fold greater levels of luciferase than H3K4b polyplexes. With gel retardation and heparin displacement assays, the H3K(+H)4b polyplexes showed greater stability than H3K4b. Intracellular uptake and co‐localization of H3K(+H)4b polyplexes within acidic endosomes were also significantly increased compared to H3K4b. Similar to H3K(+H)4b, several HK analogues with an additional histidine in the second domain of their branches were effective carriers of mRNA. When combined with DOTAP liposomes, H3K(+H)4b was synergistic in delivery of mRNA. H3K(+H)4b was a more effective carrier of mRNA than H3K4b. Mechanistic studies suggest that H3K(+H)4b polyplexes were more stable than H3K4b polyplexes. Lipopolyplexes formed with H3K(+H)4b markedly increased mRNA transfection. Branched HK mRNA polyplexes with one additional histidine in a specific location show marked improvement in their cellular uptake, accumulation within endosomes, as well as their transfection efficiency. Those HK carriers lacking an extra histidine, which form mRNA polyplexes, have decreased stability, resulting in a low transfection.
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影响因子：
9.5
作者：
Chen G;Ma B;Wang Y;Gong S
通讯作者：
Gong S
可调节的聚（胺二聚体）聚合物可调性，用于定制的核酸递送和其他生物医学应用。
影响因子：
6.2
作者：
Kauffman AC;Piotrowski-Daspit AS;Nakazawa KH;Jiang Y;Datye A;Saltzman WM
通讯作者：
Saltzman WM
影响因子：
10.8
作者：
Le Moignic, A.;Malard, V;Mateo, V
通讯作者：
Mateo, V
影响因子：
4.7
作者：
Chang, Kai-Ling;Higuchi, Yuriko;Hashida, Mitsuru
通讯作者：
Hashida, Mitsuru
DOI：
10.1016/j.nano.2013.07.008
发表时间：
2014-01-01
影响因子：
5.4
作者：
Laechelt, Ulrich;Kos, Petra;Wagner, Ernst
通讯作者：
Wagner, Ernst