Key Laboratory of Colloid and Interface Chemistry & Key Laboratory of Special Aggregated Materials, Shandong University, Ministry of Education, Jinan 250100, China.
Soft Matter. 2016 Sep 28;12(36):7495-504. doi: 10.1039/c6sm00704j. Epub 2016 Aug 19.
The thermo-reversible capture and release of DNA were studied by the protonation and deprotonation of alkyldimethylamine oxide (CnDMAO, n = 10, 12 and 14) in Tris-HCl buffer solution. DNA/C14DMAO in Tris-HCl buffer solution with pH = 7.2 is transparent at 25 °C, indicating that DNA molecules exist mainly in individuals and the binding of C14DMAO is weak. With the increase of temperature, the pH of the buffer solution continuously decreases, which leads to protonation of C14DMAO (C14DMAO + H(+)→ C14DMAOH(+)) and an obvious increase of the turbidity of the samples. This indicates a stronger binding of the protonated C14DMAOH(+) to DNA. Further investigations demonstrated the formation of DNA/C14DMAOH(+) complexes, in which the stretched DNA molecules are effectively compacted as evidenced from UV-vis absorptions, circular dichroism (CD) measurements, atomic force microscopy (AFM) observations, dynamic light scattering (DLS) measurements and agarose gel electrophoresis (AGE). Interestingly, when the temperature is turned back to 25 °C, the compacted DNA molecules can fully recover to the stretched conformation. This cycle can be repeated several times without obvious loss of efficiency. The effect of the chain length of CnDMAO has also been investigated. When C14DMAO was replaced by C12DMAO, similar phenomena can be observed with a slightly higher critical surfactant concentration for DNA compaction and a slightly lower pH of Tris-HCl buffer solution with pH = 6.8. For the DNA/C10DMAO system, however, no DNA compaction was observed even in Tris-HCl buffer solution with a much lower pH and a much higher C10DMAO concentration. The negative charges of DNA molecules can easily be neutralized by positive charges of cationic CnDMAOH(+) (n = 12 and 14) micelles. DNA was compacted and then insoluble DNA/CnDMAOH(+) complexes were formed. Because of the much higher critical micelle concentration (cmc) of the shorter chain length C10DMAOH(+), cationic C10DMAOH(+) micelles cannot form under the studied condition to compact DNA. The strategy may provide an efficient and alternative approach for stimuli-responsive gene therapy and drug release.
在 Tris-HCl 缓冲溶液中,通过烷基二甲基氧化胺(CnDMAO,n = 10、12 和 14)的质子化和去质子化来研究 DNA 的热可逆捕获和释放。在 pH = 7.2 的 Tris-HCl 缓冲溶液中,DNA/C14DMAO 在 25°C 下是透明的,这表明 DNA 分子主要以单体形式存在,并且 C14DMAO 的结合较弱。随着温度的升高,缓冲溶液的 pH 值不断降低,导致 C14DMAO 的质子化(C14DMAO + H(+)→ C14DMAOH(+))和样品浊度的明显增加。这表明质子化的 C14DMAOH(+)与 DNA 的结合更强。进一步的研究表明,形成了 DNA/C14DMAOH(+)复合物,其中伸展的 DNA 分子有效地被压缩,这可以从紫外可见吸收、圆二色性(CD)测量、原子力显微镜(AFM)观察、动态光散射(DLS)测量和琼脂糖凝胶电泳(AGE)得到证明。有趣的是,当温度恢复到 25°C 时,压缩的 DNA 分子可以完全恢复到伸展的构象。这个循环可以重复多次,而没有明显的效率损失。还研究了 CnDMAO 的链长的影响。当 C14DMAO 被 C12DMAO 取代时,在 DNA 压缩的临界表面活性剂浓度略高和 Tris-HCl 缓冲溶液的 pH 值略低(pH = 6.8)的情况下,可以观察到类似的现象。然而,对于 DNA/C10DMAO 体系,即使在 pH 值更低且 C10DMAO 浓度更高的 Tris-HCl 缓冲溶液中,也没有观察到 DNA 压缩。DNA 分子的负电荷很容易被阳离子 CnDMAOH(+)(n = 12 和 14)胶束的正电荷中和。DNA 被压缩,然后形成不溶性的 DNA/CnDMAOH(+)复合物。由于较短链长 C10DMAOH(+)的临界胶束浓度(cmc)高得多,在研究条件下,阳离子 C10DMAOH(+)胶束不能形成以压缩 DNA。该策略可能为刺激响应性基因治疗和药物释放提供一种有效且替代的方法。