利用DNA三维纳米结构对量子点功能化的有序调控

doi:10.7503/cjcu20170539

摘要/Abstract

摘要：

利用DNA纳米技术构建了内部具有空穴的DNA纳米立方体结构, 将量子点封装在其内部, 可达到在量子点的特定位点修饰数量可控的不同DNA序列的目的, 进而精准控制量子点的结合位点数量和空间取向. 为了验证构建的结构表面可以功能化不同的DNA序列, 且可控地连接在不同位点, 继续通过DNA之间的杂交对此结构进行了不同尺寸金纳米粒子的组装. 通过透射电子显微镜观察发现, 在此方法下由DNA三维纳米结构与量子点组建的复合结构不仅能控制连接的金纳米粒子数量, 还能控制组装后的几何构型. 本文方法适于构建多结合位点与功能化的量子点探针, 在生物医学方面有巨大的应用潜力.

关键词: DNA, 量子点, 功能化, 金纳米粒子

Abstract:

A fundamental unsolved problem for quantum dot(QD) chemistry is to simultaneously control the type, number and spatial orientation of QD valencies(i.e., the binding sites on each QD). Herein, we demonstrate a strategy to generate structurally predicable multivalent QD with uniform valency type, number and spatial orientation by caging the QD in a DNA cube by DNA nanotechnology. The results show that diffe-rent DNA valencies could be anchored to specific positions of the cube to form homogeneous DNA-functiona-lized QD with adjacent or diagonal valencies. As a proof-of-concept study, different sizes of gold nanoparticles(GNPs) were conjugated to each QD valency through DNA hybridization. The resulting bivalent and trivalent QD conjugates not only possess the same GNP number but also uniform assembly geometry by low magnification and high-resolution TEM. This approach holds great potential for generating multivalent and multifunc-tional QD probes for advanced biological applications.

Key words: DNA, Quantum dot, Functionalization, Gold nanoparticles

中图分类号:

O657

TrendMD:

王莉, 李智, 沈晓琴, 马楠. 利用DNA三维纳米结构对量子点功能化的有序调控. 高等学校化学学报, 2018, 39(1): 32.

WANG Li, LI Zhi, SHEN Xiaoqin, MA Nan. Programming Single Quantum Dot Valencies via DNA Caging^†. Chem. J. Chinese Universities, 2018, 39(1): 32.

图/表 12

Table 1 DNA sequences

Name	DNA sequence(5'-3')
DNA1	TCGCTGAGTAATCGTAACTCGCCTACTGGTCAGCAAGTGTGGGCACGCACACAGTAGTAATACCAGAT-GGAGTACACAAATCTG
DNA2	CTATCGGTAGAGCAGCTATGGATCCGACGTTATACTCAGCGACAGATTTGTGAGATGACCTCTGATTA-CGCGTACAACTAGCGG
DNA3	CACTGGTCAGATACAGAATCACTGGAACAGTACTACCGATAGCCGCTAGTTGAGATGCCTAAGTA-GCGCCATGAGGTTTGCTGA
DNA4	CCACACTTGCAAGTCCTAGACGGTAGCTCGAACTGACCAGTGTCAGCAAACCAGACTA-TCGATGGCATAATCCAGTGTGCGTGC
DNA5	GTACGCGTCTAGGATCAAAAAGTTCCAGTGATTCTGTA
DNA6	TCGAGCTACCGTCTAGGACT
DNA7	GACCAGTAGGCGAGTTACGA
DNA8	AACGTCGGATCCATAGCTGC
DNA9	GGATTATGCCATCGATAGTC
DNA10	CATGGCGCTACTTAGGCATC
DNA11	ACGCGTAATCAGAGGTCATC
DNA12	ACTCCATCTGGTATTACTACAAAAAAAAAAAA
DNA13(ps-po DNA)	GATCCTAGACGCGTACAAAAAG^G^G^G^G^G^G^G^G^G^TTTTTTTTTTTT
DNA14(thiolated)	CATTGATCCGAGCCTAAAAAAAAAA
DNA15(thiolated)	TGATTCCAGGTAGCAAAAAAAAAAA
DNA16(thiolated)	CGTGCTAAGTGCGATAAAAAAAAAA
DNA6'	TGCTACCTGGAATCATCGAGCTACCGTCTAGGACT
DNA7'	AGGCTCGGATCAATGGACCAGTAGGCGAGTTACGA
DNA11'	ATCGCACTTAGCACGACGCGTAATCAGAGGTCATC
DNA12'(Cy-5)	ACTCCATCTGGTATTACTACAAAAAAAAAAAA-Cy5

Scheme 1 Design of DNA cube(A), DNA cube-QDs and QDs-GNPs(B)

Table 2 Reactant volumes of gold nanoparticles with different sizes

Size/nm	V(H₂O)/mL	V(HAuCl₄)/μL	V(Citrate acid)/mL
15	25	250	1.25
20	30	300	1.05

Fig.1 5% Native PAGE characterization of DNA cubeThe expected hybridization product of each sample is illustrated above.

Fig.2 AFM image(A) and height statistics(B) of DNA cube

Fig.3 Absorption(a) and photoluminescence(b) spectra of DNA13-QD(A), native PAGE characterization of hybridization between DNA13-QD(lane 1) and DNA5(lane 2)(B) and low(C) and high-resolution(D) TEM images of DNA13-QD

Scheme 2 Schematic illustration of the assembly route of the DNA-caged QD The complementary ssDNA domains are highlighted in same colors.

Fig.4 Native PAGE characterization of heterobivalent QD(lane 1), DNA cube-QD(lane 2), and DNA cube(lane 3), low magnification(B1) and high-resolution(B2) TEM images of DNA cube-QD stained with phosphotungstic acid and AFM image(C) and height statistics(D) of DNA cube-QDThe DNA cube was pre-stained with GelRed before loading on the gel.

Fig.5 Fluorescence spectra of DNA cube-QD(a), DNA cube-QD with Cy5 labeling(b) and empty DNA cube with Cy5 labeling(c)(A) and QD-Cy5(a), QD(b), QD-Cy5+cDNA(c)(B)All samples were excited at 405 nm.

Fig.6 Schematic illustration of DNA-caged QD with adjacent(1 and 2) and diagonal(2 and 3) valencies(A), low and high magnification TEM images of GNPs assembled to bivalent DNA-caged QD with adjacent valencies(B) and diagonal valencies(C)

Fig.7 Statistics of the separation distance(center to center, inset) of GNPs conjugated to bivalent QDs with adjacent(A) and diagonal valencies(B)50 complex structures were examined in each case.

Fig.8 Assembly of different sized GNPs with DNA-caged QDs(A) Low and high magnification TEM images of small and large GNPs assembled to bivalent DNA-caged QD with adjacent position; (B) low and high magnification TEM images of small, medium and large GNPs assembled to trivalent DNA-caged QD with adjacent and diagonal position.

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