A Synthetic Platform for Non-Antibiotic Eradication of Bacterial Infections

抗生素的发现改变了医学，但抗生素的数量不断增加， 耐药细菌感染正在成为一个令人担忧的威胁，使我们需要新的抗菌剂 方法.开发新的经典抗生素可能是昂贵和耗时的， 可以快速克服这些挑战。在这项工作中，我们建议开创一个非抗生素平台， 通过改造抗微生物细菌来治疗细菌感染， 传递对致病细菌特异的有毒基因。如果这种方法成功， 将促进以最低成本快速开发新的抗菌疗法。治疗 活工程菌的细菌感染可能被认为是非正统的， 这种方法已经在概念上用于对人群进行疫苗接种， 这些疾病是由路易斯·巴斯德首先证明的。 细菌的性别是混杂的，可以在广泛的细菌中传递基因， 从供体细菌到受体细菌通过性菌毛形成更结构化的生物膜。的有毒 作为我们抗菌平台的一部分，我们设想将细菌免疫CRISPR/Cas 系统，一种基于RNA的核酸内切酶保护机制。虽然CRISPR通常用于 保护细菌免受入侵的DNA，它已被证明是致命的宿主细菌，如果 针对自己的DNA。CRISPR系统是一种令人兴奋的推定抗菌剂， 核酸酶可以靶向仅在病原菌中发现的非常特异的DNA SNP。 因此，工程化细菌携带编码靶向Cas核酸酶的接合质粒， 与经典抗生素相比，所需的病原体SNP使微生物组的其余部分保持完整。 有了这个奖项，我们建议开发这种新型的抗菌平台， 具有不同工程化细菌接合子的CRISPR/Cas系统的遗传元件 在一个无害的细菌载体的装置。我们将测试这种抗菌策略， 和体外针对多种细菌病原体的活力测定，并将这些测试转化为 动物模型系统。最后，我们将制定一个可控的战略，以避免可能不必要的 我们的工程菌在野外的繁殖。在未来，这项工作可能会导致 开发一个全新的生物医学领域，彻底改变 细菌性疾病

The discovery of antibiotics has transformed medicine, but the rising number of antibiotic resistant bacterial infections is becoming an alarming threat, leaving us in need of new antimicrobial methods. Development of new classical antibiotics can be costly and time consuming and bacteria can overcome these challenges quickly. In this work, we propose to pioneer a non-antibiotic platform to treat bacterial infections by engineering antimicrobial bacteria, in which we highjack bacterial sex to deliver toxic genes specific to disease causing pathogenic bacteria. If this approach is successful, it will facilitate the rapid development of new antimicrobial therapies at minimal cost. The treatment of bacterial infections with live engineered bacteria may be considered unorthodox however this kind of approach is already conceptually in use in the vaccination of the human population against deadly diseases as first demonstrated by Louis Pasteur. Bacterial sex is promiscuous and can deliver genes in a wide range of bacteria from planktonic to more structured biofilm forms via a sex pilus from donor to recipient bacterium. For the toxic component of our antimicrobial platform, we envision redirecting the bacterial immunity CRISPR/Cas system, an RNA based endonuclease protective mechanism. While CRISPR normally serves to protect the bacterium from invading DNA, it has been shown to be lethal to the host bacterium if targeted towards its own DNA. The CRISPR system is an exciting putative antimicrobial as the Cas nuclease can be targeted towards very specific SNPs of DNA only found in pathogenic bacteria. Thus, the engineered bacterium carries a conjugative plasmid encoding the Cas nuclease targeted to a desired pathogen SNP leaving the rest of the Microbiome intact in contrast to classical antibiotics. With this award, we propose to develop this novel antimicrobial platform combining the toxic genetic elements of the CRISPR/Cas system with different engineered bacterial conjugative apparatus in a harmless bacterial carrier. We will test this antimicrobial strategy by performing mating and viability assays against a variety of bacterial pathogens in vitro and translate these tests to animal model systems. Finally we will develop a controllable strategy to avoid possible unwanted propagation of our engineered bacteria into the wild. In the future, this work may lead to the development of a completely new field of biotherapeutic medicine and revolutionize treatment of bacterial diseases.