Antibacterial nanopores composed of DNA

DNA组成的抗菌纳米孔

• 批准号: 1764872
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1764872
• 关键词: Antibacterial; nanopores; composed; DNA

Strategic Research Priority: Industrial biotechnology and bioenergyDue to the alarming rise in antibiotic resistance and decline of active compounds, the development of new powerful bacteria-killing agents is of great importance. In this project, we will create and test bilayer-puncturing nanopores that lyse bacteria but leave eukaryotic cells unharmed. The nanopore will be composed of folded DNA and carry lipid anchors for membrane insertion. The synthetic nanopores will be characterized and examined to unterstand their selective interaction with bacterial but not human cell membranes. Synthetic DNA nanopores that mimic biological behaviour and insert into membranes have been prepared previously and attracted considerable scientific interest. But the use of synthetic pores to selectively kill bacteria is a completely new observation. The project falls with the BBSRC's strategic priorities and covers topics of synthetic biology, nanobiotechnology, chemcial biology, microbiology, and biophysics. By developing novel antibacterial compounds.The project is experimentally feasible in the time frame as it is supported by strong preliminary data generated jointly by the PI's group at UCL and the non-academic partner MR at NPL. The data show that DNA pores of 7 nm height and 5 nm width are active against both Gram-positive and Gram-negative pathogenic bacteria. Remarkably, activity is selective against bacteria but not human cells. While the findings are scientifically striking and of clear biological impact, there are several fundamental questions that have to be answered within the project in order to clarify the science and help exploit it:(i) What are the structural and chemical components of the pore that are essential for cell killing? The data suggest that the structure of a pore AND lipid anchors are essential. But can a minimal pore be constructed with the same killing activity? What is the influence of the number and position of lipid anchors on activity?(ii) What is the mechanism of bacterial cell killing? Do the pores puncture an otherwise intact membrane or completely rupture it? What are the kinetics of the membrane binding and bacterial killing? (iii) What is the reason for the selectivity of the pores? Do they recognise membrane components of the bacterial membrane which are not present in eucaryotic cells?(iv) Can bacteria-killing DNA pores be developed further and serve as conceptual template to create related nanomaterials or organic small-molecule drugs that achieve selective killing?The proposed project is relevant to the non-academic partner, who has a long-standing and strategic interest in developing advanced measurement approaches, materials and methods to address the problem of antimicrobial resistance.The elibility of the non-academic partner NPL has been confirmed with Dr. Nadine Mogford.The projects is supported by a strong track-record of the SH and MR in the relevant areas (see point 4 below) and benefits from previous joint collaboration between SH and MR in the supervision of a joint PhD student.

战略研究重点：工业生物技术和生物能源由于抗生素耐药性的惊人上升和活性化合物的下降，开发新型强效杀菌剂非常重要。在这个项目中，我们将创建并测试双层穿刺纳米孔，它可以裂解细菌，但不会伤害真核细胞。纳米孔将由折叠的 DNA 组成，并带有用于膜插入的脂质锚。将对合成纳米孔进行表征和检查，以了解它们与细菌而不是人类细胞膜的选择性相互作用。先前已经制备了模仿生物行为并插入膜中的合成 DNA 纳米孔，并引起了相当大的科学兴趣。但利用合成孔隙来选择性杀死细菌是一个全新的观察结果。该项目符合 BBSRC 的战略重点，涵盖合成生物学、纳米生物技术、化学生物学、微生物学和生物物理学等主题。通过开发新型抗菌化合物。该项目在时间范围内在实验上是可行的，因为它得到了伦敦大学学院的 PI 小组和 NPL 的非学术合作伙伴 MR 联合生成的强有力的初步数据的支持。数据显示，高度为 7 nm、宽度为 5 nm 的 DNA 孔对革兰氏阳性和革兰氏阴性病原菌均具有活性。值得注意的是，该活性对细菌有选择性，但对人体细胞没有选择性。虽然这些发现在科学上令人震惊并且具有明显的生物学影响，但为了阐明科学并帮助利用它，该项目中必须回答几个基本问​​题：（i）对于杀死细胞至关重要的孔的结构和化学成分是什么？数据表明孔和脂质锚的结构至关重要。但能否构建出具有相同杀伤活性的最小孔呢？脂质锚的数量和位置对活性有何影响？(ii) 杀灭细菌细胞的机制是什么？这些孔是否会刺穿原本完好的膜或使其完全破裂？膜结合和细菌杀灭的动力学是什么？ (iii) 孔隙选择性的原因是什么？它们是否能识别真核细胞中不存在的细菌膜成分？（iv）是否可以进一步开发杀菌DNA孔，并作为概念模板来创建相关纳米材料或有机小分子药物，实现选择性杀灭？拟议项目与非学术合作伙伴相关，他们对开发先进的测量方法、材料和方法具有长期和战略兴趣 解决抗菌素耐药性问题。非学术合作伙伴 NPL 的资格已得到 Nadine Mogford 博士的确认。这些项目得到了 SH 和 MR 在相关领域的良好记录的支持（见下文第 4 点），并受益于 SH 和 MR 之前在联合博士生的监督下的联合合作。