Exploiting the Protein Corona from Cell Lysate on DNA Functionalized Gold Nanoparticles for Enhanced mRNA Translation
Exploiting the Protein Corona from Cell Lysate on DNA Functionalized Gold Nanoparticles for Enhanced mRNA Translation
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DOI：
10.1021/acsami.6b15269
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发表时间：
2017-03-29
影响因子：
9.5
通讯作者：
Kah, James Chen Yong
中科院分区：
文献类型：
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作者：
Chan, Kian Ping;Gao, Yang;Kah, James Chen Yong
这项研究描述了使用DNA功能化的金纳米颗粒(AuNPs)来增强细胞裂解物中蛋白质的合成，并研究了增强mRNA翻译的机制。通过与胰岛素和绿色荧光蛋白(GFP)这两个感兴趣的mRNA的3‘-非翻译区杂交的合适的DNA寡聚体序列，我们发现这些DNA结合的AuNPs(AuNP-DNA)被导入HeLa细胞裂解物中，分别比仅有mRNA的基线产量增加2.18倍和1.80倍。非DNA柠檬酸盐封端的AuNP(1.25倍)和带有非特异性PolyT(T)序列的AuNP-DNA(1.25倍)显著减少胰岛素合成。我们发现，核糖体的非特异性吸附和翻译因子在AuNP-DNA上形成裂解蛋白冠状结构以及DNA寡聚体与感兴趣的mRNA之间的弱杂交都是导致翻译因子、核糖体和mRNA紧密结合的重要因素。这可以减少核糖体在mRNA翻译过程中的循环时间，从而提高蛋白质合成的效率。这项工作的结果表明，通过合理的DNA设计，有可能用AuNP-DNA来调节细胞内的生物过程，并增加它们在各种生物医学应用中的蛋白质产量。
This study describes the use of DNA functionalized gold nanoparticles (AuNPs) to enhance the synthesis of proteins in cell lysate and examines the mechanisms behind the enhanced mRNA translation. With an appropriate DNA oligomer sequence that hybridizes to the 3 '-untranslated region of two mRNA of interest, insulin and green fluorescent protein (GFP), we found that these DNA conjugated AuNPs (AuNP-DNA) introduced into HeLa cell lysate enhanced the synthesis of insulin and GFP by up to 2.18 and 1.80-fold, respectively, over baseline production with just the mRNA present. The insulin synthesis was markedly reduced with non-DNA citrate-capped AuNP (1.25-fold) and AuNP-DNA with a nonspecific poly(T) sequence (1.25-fold). We showed that both nonspecific adsorption of ribosomes and translation factors to form a lysate protein corona on AuNP-DNA and weak hybridization between DNA oligomers and mRNA of interest were important factors that brought translation factors, ribosomes, and mRNA into close proximity of each other. This could reduce the recycling time of ribosomes during mRNA translation, thereby increasing the efficiency of protein synthesis. The outcome of this work shows that with rational DNA design, it could be possible to modulate intracellular biological processes with AuNP-DNA and increase their production of proteins for various biomedical applications.