Antibacterial nanopores composed of DNA

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，开发新的强效杀菌剂至关重要。在这个项目中，我们将创建和测试双层穿刺纳米孔，裂解细菌，但不伤害真核细胞。纳米孔将由折叠的DNA组成，并携带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。先前已经制备了模拟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。但是使用合成孔选择性地杀死细菌是一个全新的观察。该项目福尔斯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的主题。通过开发新型抗菌化合物。该项目在实验上是可行的，因为它得到了UCL PI团队和NPL非学术合作伙伴MR共同生成的强大初步数据的支持。数据显示，7 nm高和5 nm宽的DNA孔对革兰氏阳性和革兰氏阴性病原菌都有活性。值得注意的是，活性对细菌有选择性，但对人类细胞没有选择性。虽然这些发现在科学上是惊人的，并且具有明显的生物学影响，但为了澄清科学并帮助利用它，该项目必须回答几个基本问题：（i）孔的结构和化学成分是什么？这些数据表明，孔和脂质锚的结构是必不可少的。但是一个最小的孔能被构造成具有同样的杀伤活性吗？脂质锚的数量和位置对活性有什么影响？(ii)细菌细胞的杀伤机制是什么？这些小孔是刺穿了一个完整的膜，还是使它完全破裂？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)杀细菌的DNA孔是否可以进一步开发，并作为概念模板来创建相关的纳米材料或实现选择性杀伤的有机小分子药物？拟议的项目与非学术合作伙伴有关，他们对开发先进的测量方法具有长期的战略兴趣，材料和方法来解决抗菌素耐药性的问题。非学术合作伙伴NPL的可行性已经得到了Nadine Mogford博士的确认。该项目得到了SH和MR在相关领域的强大跟踪记录的支持（见下文第4点），并受益于SH和MR之前在联合博士生监督下的联合合作。

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.