Departments of Bioengineering, Materials Science and Engineering and Beckman Institute, Mills Breast Cancer Institute, and Carle Foundation Hospital, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
Department of Biochemistry, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801, United States.
ACS Appl Mater Interfaces. 2020 Apr 8;12(14):16137-16149. doi: 10.1021/acsami.0c02463. Epub 2020 Mar 26.
By using complementary DNA sequences as surface ligands, we selectively allow two individual diffusing "dual-color" carbon dots to interact and . Spontaneous nanoscale oxidation of surface-abundant nitroso-/nitro-functionalities leads to two distinctly colored carbon dots (CD) which are isolated by polarity driven chromatographic separation. Green- and red-emitting carbon dots (gCD and rCD) were decorated by complementary single-stranded DNAs which produce a marked increase in the fluorescence emission of the respective carbon dots. Mutual colloidal interactions are achieved through hybridization of complementary DNA base pairs attached to the respective particles, resulting in quenching of their photoluminescence. The observed post-hybridization quenching is presumably due to a combined effect from an aggregation of CDs post duplex DNA formation and close proximity of multicolored CDs, having overlapped spectral regions leading to a nonradiative energy transfer process possibly released as heat. This strategy may contribute to the rational design of mutually interacting carbon dots for a better control over the resulting assembly structure for studying different biological phenomenon including molecular cytogenetics. One of the newly synthesized CDs was successfully used to image intracellular location of GAPDH mRNA using an event of change in fluorescence intensity (FI) of CDs. This selectivity was introduced by conjugating an oligonucleotide harboring complementary sequence to GAPDH mRNA. FI of this conjugated carbon dot, rCD-GAPDH, was also found to decrease in the presence of Ca, varied in relation to H concentrations, and could serve as a tool to quantify the intracellular concentrations of Ca and pH value (H) which can give important information about cell survival. Therefore, CD-oligonucleotide conjugates could serve as efficient probes for cellular events and interventions.
通过使用互补 DNA 序列作为表面配体,我们选择性地允许两个单独扩散的“双色”碳点相互作用和结合。表面丰富的亚硝基/硝基官能团的自发纳米尺度氧化导致两个明显不同颜色的碳点(CD),通过极性驱动的色谱分离将其分离。绿色和红色发射的碳点(gCD 和 rCD)被互补单链 DNA 修饰,这导致各自碳点的荧光发射明显增加。通过附着在各自颗粒上的互补 DNA 碱基对的杂交实现了相互胶体相互作用,导致其光致发光猝灭。观察到的杂交后猝灭可能是由于形成双链 DNA 后 CD 的聚集和多色 CD 的紧密接近的综合效应,这导致重叠光谱区域的非辐射能量转移过程可能以热的形式释放。该策略可能有助于合理设计相互作用的碳点,以更好地控制组装结构,从而研究包括分子细胞遗传学在内的不同生物学现象。新合成的 CD 之一成功地用于使用 CD 荧光强度(FI)变化的事件来对 GAPDH mRNA 的细胞内位置进行成像。这种选择性是通过将含有互补序列的寡核苷酸与 GAPDH mRNA 缀合引入的。还发现这种缀合的碳点 rCD-GAPDH 的 FI 在 Ca 的存在下降低,与 H 浓度有关,并且可以用作定量细胞内 Ca 和 pH 值(H)浓度的工具,这可以提供有关细胞存活的重要信息。因此,CD-寡核苷酸缀合物可以作为细胞事件和干预的有效探针。