Targeted intracellular delivery of proteins with spatial and temporal control.
Targeted intracellular delivery of proteins with spatial and temporal control.
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DOI：
10.1021/mp500675p
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发表时间：
2015-02-02
影响因子：
4.9
通讯作者：
Reich NO
中科院分区：
文献类型：
--
作者：
Morales DP;Braun GB;Pallaoro A;Chen R;Huang X;Zasadzinski JA;Reich NO
虽然存在许多将遗传物质或小分子递送到细胞的方法，但很少有可用于将蛋白质递送到胞质溶胶的方法。我们描述了一种模块化的光激活纳米载体，其通过受体介导的内吞作用将蛋白质转运到细胞中，并通过光触发的内体逃逸将货物递送到胞质溶胶。该平台基于中空金纳米壳（HGN），其具有通过亲合力增强的镍螯合连接层连接的多组氨酸标记的蛋白质;在这里，我们使用绿色荧光蛋白（GFP）作为模型可交付货物。载有GFP的纳米载体的内体摄取由与GFP融合的C-末端规则（CendR）内化肽介导。聚焦飞秒脉冲激光激发触发蛋白质从纳米载体释放和内体破坏，并且释放的蛋白质能够靶向核仁，一种模型细胞内细胞器。我们通过加载和释放Sox 2和p53进一步证明了该方法的通用性。这种用于靶向单个细胞的方法，具有类似于显微注射的分辨率，提供了对蛋白质递送的空间和时间控制。
While a host of methods exist to deliver genetic materials or small molecules to cells, very few are available for protein delivery to the cytosol. We describe a modular, light-activated nanocarrier that transports proteins into cells by receptor-mediated endocytosis and delivers the cargo to the cytosol by light triggered endosomal escape. The platform is based on hollow gold nanoshells (HGN) with polyhistidine tagged proteins attached through an avidity-enhanced, nickel chelation linking layer; here, we used green fluorescent protein (GFP) as a model deliverable cargo. Endosomal uptake of the GFP loaded nanocarrier was mediated by a C-end Rule (CendR) internalizing peptide fused to the GFP. Focused femtosecond pulsed-laser excitation triggered protein release from the nanocarrier and endosome disruption, and the released protein was capable of targeting the nucleoli, a model intracellular organelle. We further demonstrate the generality of the approach by loading and releasing Sox2 and p53. This method for targeting of individual cells, with resolution similar to microinjection, provides spatial and temporal control over protein delivery.
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影响因子：
17.1
作者：
Boeneman, Kelly;Delehanty, James B.;Blanco-Canosa, Juan B.;Susumu, Kimihiro;Stewart, Michael H.;Oh, Eunkeu;Huston, Alan L.;Dawson, Glyn;Ingale, Sampat;Walters, Ryan;Domowicz, Miriam;Deschamps, Jeffrey R.;Algar, W. Russ;DiMaggio, Stassi;Manono, Janet;Spillmann, Christopher M.;Thompson, Darren;Jennings, Travis L.;Dawson, Philip E.;Medintz, Igor L.
通讯作者：
Medintz, Igor L.
影响因子：
10.8
作者：
Gu, Zhen;Yan, Ming;Tang, Yi
通讯作者：
Tang, Yi
影响因子：
64.8
作者：
INGRAM, VM
通讯作者：
INGRAM, VM
影响因子：
15
作者：
Chen, CC;Lin, YP;Wu, YC
通讯作者：
Wu, YC
影响因子：
4.9
作者：
Jana, NR;Gearheart, L;Murphy, CJ
通讯作者：
Murphy, CJ