P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri.
P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri.
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DOI：
10.1021/acssynbio.2c00465
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发表时间：
2023-03-17
影响因子：
4.7
通讯作者：
Wang, Baojun
中科院分区：
文献类型：
--
作者：
Huan, Yang W.;Torraca, Vincenzo;Brown, Russell;Fa-arun, Jidapha;Miles, Sydney L.;Oyarzun, Diego A.;Mostowy, Serge;Wang, Baojun
成簇的、规则间隔的短回文重复序列（CRISPR）和Cas9 RNA引导的核酸酶的发现为选择性杀死特定细菌种群或物种提供了前所未有的机会。然而，使用CRISPR-Cas9清除体内细菌感染受到cas9基因构建体向细菌细胞中的低效递送的阻碍。在这里，我们使用一种广泛的宿主范围P1衍生的噬菌粒将CRISPR-Cas9染色体靶向系统递送到大肠杆菌和致炎性福氏志贺菌中，以实现靶向细菌细胞的DNA序列特异性杀伤。我们表明，辅助P1噬菌体DNA包装位点（pac）的遗传修饰显著提高了包装的噬菌粒的纯度，并改善了Cas9介导的对S.弗氏细胞我们进一步证明了P1噬菌体颗粒可以将染色体靶向cas9噬菌粒递送到S. flexnerin体内使用斑马鱼幼虫感染模型，其中它们显著降低细菌负荷并促进宿主存活。我们的研究强调了将基于P1噬菌体的递送与CRISPR染色体靶向系统相结合的潜力，以实现DNA序列特异性细胞杀伤力和细菌感染的有效清除。
The discovery of clustered, regularly interspaced, short palindromic repeats (CRISPR) and the Cas9 RNA-guided nuclease provides unprecedented opportunities to selectively kill specific populations or species of bacteria. However, the use of CRISPR-Cas9 to clear bacterial infections in vivo is hampered by the inefficient delivery of cas9 genetic constructs into bacterial cells. Here, we use a broad-host-range P1-derived phagemid to deliver the CRISPR-Cas9 chromosomal-targeting system into Escherichia coli and the dysentery-causing Shigella flexneri to achieve DNA sequence-specific killing of targeted bacterial cells. We show that genetic modification of the helper P1 phage DNA packaging site (pac) significantly enhances the purity of packaged phagemid and improves the Cas9-mediated killing of S. flexneri cells. We further demonstrate that P1 phage particles can deliver chromosomal-targeting cas9 phagemids into S. flexneriin vivo using a zebrafish larvae infection model, where they significantly reduce the bacterial load and promote host survival. Our study highlights the potential of combining P1 bacteriophage-based delivery with the CRISPR chromosomal-targeting system to achieve DNA sequence-specific cell lethality and efficient clearance of bacterial infection.
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DOI：
10.3390/v14040678
发表时间：
2022-03-25
期刊：
Viruses
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.7
作者：
IIDA, S;STREIFF, MB;ARBER, W
通讯作者：
ARBER, W
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：
30.3
作者：
Depardieu, Florence;Didier, Jean-Philippe;Bikard, David
通讯作者：
Bikard, David
影响因子：
168.9
作者：
Kotloff, Karen L.;Nataro, James P.;Levine, Myron M.
通讯作者：
Levine, Myron M.