Removal of AMR plasmids using a mobile, broad host-range CRISPR-Cas9 delivery tool.
Removal of AMR plasmids using a mobile, broad host-range CRISPR-Cas9 delivery tool.
复制标题
DOI：
10.1099/mic.0.001334
复制
发表时间：
2023-05
期刊：
影响因子：
2.8
通讯作者：
van Houte, Stineke
中科院分区：
文献类型：
--
作者：
Walker-Sunderhauf, David;Klumper, Uli;Pursey, Elizabeth;Westra, Edze R.;Gaze, William H.;van Houte, Stineke
关键词：
开发了一种广泛宿主范围的CRISPR-Cas9递送工具，用于接合去除抗菌素耐药(AMR)质粒(TOP)，并保护一系列大肠埃希菌和假单胞菌免受AMR质粒的摄取(LOTOR)。抗菌素耐药(AMR)基因广泛分布于质粒上。因此，旨在阻止质粒摄取和转移的干预措施可能会抑制AMR的传播。以前的研究使用了基于CRISPR-Cas的技术来从目标细菌中去除编码AMR基因的质粒，使用基于噬菌体或质粒的递送载体，这些载体通常具有较窄的宿主范围。为了使这项技术用于从复杂微生物群落的多个成员中去除AMR质粒是可行的，需要一种高效、广泛的宿主范围递送工具。我们设计了广泛宿主范围的IncP1-质粒pKJK5来编码针对AMR基因的Cas9。我们证明了得到的pKJK5：：CSG有能力阻止AMR质粒的摄取，并从大肠杆菌中去除驻留的质粒。此外，由于其广泛的宿主范围，pKJK5：：CSG成功地阻止了一系列环境、猪和人相关的大肠埃希菌以及两种假单胞菌的AMR质粒的摄取。这项研究确立了pKJK5：：CSG作为一种有前景的广泛宿主范围的CRISPR-Cas9载体工具来去除AMR质粒，它有可能应用于复杂的微生物群落，从广泛的细菌物种中去除AMR基因。
A broad host-range CRISPR-Cas9 delivery tool was developed and used to conjugatively remove antimicrobial resistance (AMR) plasmids (top) and protect a series of coliform and Pseudomonas isolates from AMR plasmid uptake (bottom). Antimicrobial resistance (AMR) genes are widely disseminated on plasmids. Therefore, interventions aimed at blocking plasmid uptake and transfer may curb the spread of AMR. Previous studies have used CRISPR-Cas-based technology to remove plasmids encoding AMR genes from target bacteria, using either phage- or plasmid-based delivery vehicles that typically have narrow host ranges. To make this technology feasible for removal of AMR plasmids from multiple members of complex microbial communities, an efficient, broad host-range delivery vehicle is needed. We engineered the broad host-range IncP1-plasmid pKJK5 to encode cas9 programmed to target an AMR gene. We demonstrate that the resulting plasmid pKJK5::csg has the ability to block the uptake of AMR plasmids and to remove resident plasmids from Escherichia coli . Furthermore, due to its broad host range, pKJK5::csg successfully blocked AMR plasmid uptake in a range of environmental, pig- and human-associated coliform isolates, as well as in isolates of two species of Pseudomonas . This study firmly establishes pKJK5::csg as a promising broad host-range CRISPR-Cas9 delivery tool for AMR plasmid removal, which has the potential to be applied in complex microbial communities to remove AMR genes from a broad range of bacterial species.
登录
查看更多内容
影响因子：
46.9
作者：
Bikard, David;Euler, Chad W.;Jiang, Wenyan;Nussenzweig, Philip M.;Goldberg, Gregory W.;Duportet, Xavier;Fischetti, Vincent A.;Marraffini, Luciano A.
通讯作者：
Marraffini, Luciano A.
影响因子：
3.5
作者：
Cormack, BP;Valdivia, RH;Falkow, S
通讯作者：
Falkow, S
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
3.2
作者：
Ferrieres, Lionel;Hemery, Gaelle;Ghigo, Jean-Marc
通讯作者：
Ghigo, Jean-Marc
影响因子：
2.6
作者：
Bahl, Martin Iain;Hansen, Lars Hestbjerg;Sorensen, Soren J.
通讯作者：
Sorensen, Soren J.