摘要
      抗生素耐药性基因的进化和传播正在引发严重的健康和环境问题。虽然环境过程是防止ARGs传播的关键障碍，但它们往往同时也是ARGs的来源，因为可能需要ARGs并在生物处理单元中积累。升级环境生物技术是当务之急。ARGs根据DNA序列而不是DNA分子的化学性质赋予抗生素耐药性。如果ARG的序列被破坏，则可以认为其已降级。因此，我们在这里提出，CRISPR-Cas免疫，一种用于消除入侵的外源DNA的古菌和细菌免疫系统，可以被重新利用并针对ARGs的降解进行定制。通过部署人工IncP机器，所设计的系统，即VADER，可以通过细菌接合成功递送。然后，我们提出了一个新的ARG降解部门，作为环境过程框架中生物单元的补充。在这项工作中，设计了一个10mL规模的原型缀合反应器，并在反应器中接受VADER的转缀合微生物中消除了100%的靶ARG。通过建立合成生物学和环境生物技术的联系，我们相信我们的工作不仅是解决ARG问题的一项事业，也是未来管理不需要的遗传物质的潜在解决方案。
Abstract
The evolution and dissemination of antibiotic resistance genes (ARGs) are prompting severe health and environmental issues. While environmental processes are key barriers preventing the spread of ARGs, they are often sources of ARGs at the same time, as ARGs may be required and accumulate in the biological treatment units. An upgrading of environmental biotechnology is imperative and urgent. ARGs confer antibiotic resistance based on the DNA sequences rather than the chemistry of DNA molecules. An ARG can be considered degraded if its sequence was disrupted. Therefore, we present here that CRISPR-Cas immunity, an archaeal and bacterial immune system for eliminating invading foreign DNAs, can be repurposed and tailored for the degradation of ARGs. By deploying an artificial IncP machinery, the designed system, namely VADER, can be successfully delivered via bacterial conjugation. Then, we propose a new sector for ARG degradation to be implemented as a complement to the biological units in the framework of environmental processes. In this endeavor, a prototype conjugation reactor at a 10-mL-scale was devised, and 100% of the target ARG were eliminated in the transconjugated microbes receiving VADER in the reactor. By generating a nexus of synthetic biology and environmental biotechnology, we believe that our work is not only an enterprise for tackling ARG problems but also a potential solution for managing undesired genetic materials in general in the future.

https://www.biorxiv.org/content/10.1101/2022.03.09.483686v2.abstract