Engineering bacteriophages for modified functionality

随着世界各地耐药性细菌感染病例的迅速增加，我们正在接受许多人现在所说的后抗生素时代的现实和挑战。在寻找新的细菌感染抑制剂的多种尝试中，噬菌体（Bacteriophage，简称BPHs）已经复苏。长期以来，这些细菌病毒一直被吹捧为抗生素的潜在替代品（在噬菌体疗法中），但它们的特性-最重要的是它们识别和杀死特定细菌的能力-也使它们适合于大量其他应用：从生物控制到细菌检测，从治疗载体到工程材料。虽然基于噬菌体的技术仍处于起步阶段，但它们具有广泛的影响力，可以影响我们生活的许多方面。 开发基于噬菌体的技术的第一步通常涉及筛选从自然环境中采样的噬菌体，并表征其特性，希望找到适合感兴趣应用的合适方法。这个过程可能很漫长，而且并不总是成功的。噬菌体工程是一种快速获得兴趣的替代方法;它包括通过基因工程和病毒体功能化修饰或组合已知噬菌体的特征和性质。这种有前途的策略目前受到可用于噬菌体的遗传工具和方法的限制，以及在噬菌体生物学，遗传学和调控方面仍然存在的知识差距。事实上，在大多数情况下，噬菌体工程缺乏完善的协议和工具，这些协议和工具极大地促进了细菌和酵母的基因工程。这严重限制了基于噬菌体的技术的新应用的开发和广度。该计划将包括三个主要目标：1）改进噬菌体表达盒的设计和性能; 2）设计噬菌体病毒体的结构修饰，以改善和控制其特性; 3）开发基于工程改造的ghost噬菌体的细菌瞬时修饰平台。 该研究计划将提供有关噬菌体工程的新知识，并加速在各种领域（包括健康，诊断，食品安全和环境）中基于噬菌体的应用的发展。

As the number of cases of infections by antibiotic-resistant bacteria rapidly increases around the World, we are coming to terms with the realities and challenges of what many now call the post-antibiotics era. Among the multiple attempts to find new deterrents to bacterial infections, bacteriophages (phages) have made a resurgence. These viruses of bacteria have long been touted has potential alternatives to antibiotics (in phage therapy) but their properties - most importantly their capacity to recognize and kill specific bacteria - also make them amenable to a plethora of other applications: from biocontrol to bacterial detection, from vectors for therapeutic delivery to engineered materials. While phage-based technologies are still in their infancy, they have broad reach and can impact many aspects of our lives. The first step in developing phage-based technologies typically involves screening for phages sampled from natural environments and characterizing their properties in the hope of finding a proper fit for the application of interest. This process can be lengthy and is not always successful. Phage engineering is an alternative approach rapidly garnering interest; it consists of modifying or combining the features and properties of known phages through genetic engineering and virion functionalization. This promising strategy is currently limited by the genetic tools and methods available for phages, and the significant gaps in knowledge that still exist with regards to phage biology, genetics and regulation. In fact, for the most part, phage engineering lacks the well-established protocols and tools that have greatly enabled genetic engineering of bacteria and yeast. This critically limits the development and breadth of novel applications for phage-based technologies. The proposed program will consist of three main objectives: 1) improve the design and performance of expression cassettes in phages; 2) design structural modifications of phage virions for the improvement and control of their properties; and 3) develop a platform for the transient modification of bacteria based on engineered ghost phages. This research program will provide new knowledge on phage engineering and accelerate the development of phage-based applications in a wide variety of fields, including health, diagnostics, food security, and the environment.