Evaluating the Mechanisms of Light-Triggered siRNA Release from Nanoshells for Temporal Control Over Gene Regulation.
Evaluating the Mechanisms of Light-Triggered siRNA Release from Nanoshells for Temporal Control Over Gene Regulation.
复制标题
DOI：
10.1021/acs.nanolett.8b00681
复制
发表时间：
2018-06-13
期刊：
影响因子：
10.8
通讯作者：
Day ES
中科院分区：
文献类型：
--
作者：
Riley RS;Dang MN;Billingsley MM;Abraham B;Gundlach L;Day ES
通过小干扰RNA(SiRNAs)等外源核酸调节细胞内基因表达的能力在改善疾病的研究和治疗方面具有巨大的潜力。然而，大多数用于细胞内核酸输送的转染剂和纳米颗粒载体无法区分疾病细胞和健康细胞，这可能导致它们产生意想不到的广泛基因调控。理想的递送系统只会对外界刺激做出反应，使病变组织中的靶向蛋白质沉默。为了能够在时空上控制基因沉默，研究人员已经开始开发核酸-纳米颗粒结合物，使其核酸货物保持不活动，直到通过外部施加的近红外激光按需从纳米颗粒中释放出来。这一策略可以克服其他核酸输送系统的几个局限性，但这些平台的运作机制仍不清楚。在这里，我们对硅核/金壳纳米壳(NSS)在脉冲或连续波(CW)近红外(NIR)光激发下释放结合的siRNA的机制进行了详细的研究，目的是深入了解这些纳米结合物如何实现按需基因调控。我们证明了脉冲激光照射下从NSS释放siRNA是一个与温度无关的过程，比由连续激光照射触发的siRNA释放效率要高得多。与文献相反，只有脉冲辐射才能释放siRNA双链，我们发现两种辐射模式都释放了siRNA双链和单链寡核苷酸的混合物，但脉冲辐射导致了更高比例的双链释放。为了证明脉冲照射后NSS释放的siRNA仍然具有功能，我们评估了包被绿色荧光蛋白(GFP)靶向siRNA(siGFP-NS)的NSS在细胞中按需敲除GFP的使用。我们发现，经siGFP-NS处理和脉冲激光照射的GFP表达细胞与未经激光处理的细胞相比，GFP表达下降了33%。此外，我们观察到由siGFP-NS介导的光触发基因沉默比使用商业转染剂更有效地将siRNA传递到细胞中。这项工作对等离子体NSS在光照射下释放siRNA的机制提供了前所未有的见解，并证明了彻底表征光响应纳米系统在触发基因调控中应用的重要性。
The ability to regulate intracellular gene expression with exogenous nucleic acids such as small interfering RNAs (siRNAs) has substantial potential to improve the study and treatment of disease. However, most transfection agents and nanoparticle-based carriers that are used for the intracellular delivery of nucleic acids cannot distinguish between diseased and healthy cells, which may cause them to yield unintended widespread gene regulation. An ideal delivery system would only silence targeted proteins in diseased tissue in response to an external stimulus. To enable spatiotemporal control over gene silencing, researchers have begun to develop nucleic acid-nanoparticle conjugates that keep their nucleic acid cargo inactive until it is released from the nanoparticle on-demand by externally applied near-infrared laser light. This strategy can overcome several limitations of other nucleic acid delivery systems, but the mechanisms by which these platforms operate remain ill understood. Here, we perform a detailed investigation of the mechanisms by which silica core/gold shell nanoshells (NSs) release conjugated siRNA upon excitation with either pulsed or continuous wave (CW) near-infrared (NIR) light, with the goal of providing insight into how these nanoconjugates can enable on-demand gene regulation. We demonstrate that siRNA release from NSs upon pulsed laser irradiation is a temperature-independent process that is substantially more efficient than siRNA release triggered by CW irradiation. Contrary to literature, which suggests that only pulsed irradiation releases siRNA duplexes, we found that both modes of irradiation release a mixture of siRNA duplexes and single-stranded oligonucleotides, but that pulsed irradiation results in a higher percentage of released duplexes. To demonstrate that the siRNA released from NSs upon pulsed irradiation remains functional, we evaluated the use of NSs coated with green fluorescent protein (GFP)-targeted siRNA (siGFP-NS) for on-demand knockdown of GFP in cells. We found that GFP-expressing cells treated with siGFP-NS and irradiated with a pulsed laser experienced a 33% decrease in GFP expression compared to cells treated with no laser. Further, we observed that light-triggered gene silencing mediated by siGFP-NS is more potent than using commercial transfection agents to deliver siRNA into cells. This work provides unprecedented insight into the mechanisms by which plasmonic NSs release siRNA upon light irradiation and demonstrates the importance of thoroughly characterizing photoresponsive nanosystems for applications in triggered gene regulation.
登录
查看更多内容
影响因子：
15
作者：
Huschka, Ryan;Zuloaga, Jorge;Knight, Mark W.;Brown, Lisa V.;Nordlander, Peter;Halas, Naomi J.
通讯作者：
Halas, Naomi J.
DOI：
10.1016/s0006-291x(02)02013-2
发表时间：
2002-08-30
影响因子：
3.1
作者：
Bertrand, JR;Pottier, M;Malvy, C
通讯作者：
Malvy, C
TAT肽官能化的金纳米级：使用超低辐照度增强了细胞内递送和有效的NIR光热治疗。
影响因子：
15
作者：
Yuan, Hsiangkuo;Fales, Andrew M.;Vo-Dinh, Tuan
通讯作者：
Vo-Dinh, Tuan
影响因子：
11.2
作者：
Lu W;Zhang G;Zhang R;Flores LG 2nd;Huang Q;Gelovani JG;Li C
通讯作者：
Li C
影响因子：
17.1
作者：
Wijaya, Andy;Schaffer, Stefan B.;Hamad-Schifferli, Kimberly
通讯作者：
Hamad-Schifferli, Kimberly