Applications of TiO2-DNA nanocomposites

TiO2-DNA纳米复合材料的应用

• 批准号: 7046727
• 负责人: GAYLE E. WOLOSCHAK
• 金额: $ 29.36万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7046727
• 关键词: Alphaherpesvirinae; DNA damage; acyclovir; antisense nucleic acid; biotechnology; cell line; chemical cleavage; drug resistance; endonuclease; enzyme activity; gene mutation; guanosine; metal oxide; nanotechnology; nucleic acid sequence; nucleoside analog; oligonucleotides; polymerase chain reaction; radiotracer; technology /technique development; thymidine kinase; titanium; virus DNA; virus RNA

DESCRIPTION (provided by applicant): The purpose of this project is to test the possibility of developing TiO2 -biopolymer nanocomposites as new vehicles for biotechnology. We propose to use TiO2-DNA nanocomposites as inducible gene specific endonucleases, with allele-differentiating sequence specificity. This function of nanocomposites relies on (a) TiO2 nanoparticle dependent charge separation inducing DNA cleavage; and (b) sequence specificity of oligonucleotides attached to TiO2 nanoparticle. The specific aims of this proposal are: 1. In vitro studies (outside cells) of the interaction between HSVtk specific nanocomposites and DNA and RNA specific for the HSVtk (Herpes Simplex Virus thymidine kinase) gene 2. Studies of the interaction between HSVtk specific nanocomposites and DNA specific for the HSVtk gene in cells in culture 3. Evaluation of mutations caused by interactions between HSVtk specific nanocomposites and HSVtk gene in cells in culture. We will study DNA scission following activation of single and multiple nanocomposites (with one or two different oligonucleotides attached per nanoparticle) annealed to the target sequence. The target sequence is Herpes simplex virus thymidine kinase gene (HSVtk). We chose this sequence to be our target because HSVtk acts as a suicide gene in cells treated with guanine analogues, and therefore mutagenesis of the HSVtk sequence provides cells with a selective advantage when guanine analogues are added into the cell medium. We believe that upon completion of these studies we will be able to link results of in vitro DNA scission assays with the spectra and frequency of mutations in cells in culture, use this knowledge to predict the behavior of nanocomposites inside cells, and associate the type of mutation that can be anticipated with different nanocomposite sequences and modes of nanocomposite activation.

描述（由申请人提供）：该项目的目的是测试开发 TiO2-生物聚合物纳米复合材料作为生物技术新载体的可能性。我们建议使用 TiO2-DNA 纳米复合材料作为诱导型基因特异性核酸内切酶，具有等位基因分化序列特异性。纳米复合材料的这一功能依赖于 (a) TiO2 纳米粒子依赖的电荷分离诱导 DNA 裂解； (b) 附着在 TiO2 纳米颗粒上的寡核苷酸的序列特异性。 该提案的具体目标是： 1. HSVtk 特异性纳米复合材料与 HSVtk（单纯疱疹病毒胸苷激酶）基因特异性 DNA 和 RNA 之间相互作用的体外研究（细胞外） 2. HSVtk特异性纳米复合材料与培养细胞中HSVtk基因特异性DNA之间相互作用的研究 3.评估HSVtk特异性纳米复合材料与培养细胞中HSVtk基因之间相互作用引起的突变。 我们将研究退火到靶序列上的单个和多个纳米复合材料（每个纳米颗粒附着一种或两种不同的寡核苷酸）激活后的 DNA 断裂。靶序列是单纯疱疹病毒胸苷激酶基因(HSVtk)。我们选择这个序列作为我们的靶标，因为HSVtk在用鸟嘌呤类似物处理的细胞中充当自杀基因，因此当将鸟嘌呤类似物添加到细胞培养基中时，HSVtk序列的诱变为细胞提供了选择优势。我们相信，完成这些研究后，我们将能够将体外 DNA 断裂测定的结果与培养细胞中突变的谱和频率联系起来，利用这些知识来预测细胞内纳米复合材料的行为，并将可以通过不同的纳米复合材料序列和纳米复合材料激活模式来预期突变。