Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY, USA.
Methods Mol Biol. 2024;2793:257-271. doi: 10.1007/978-1-0716-3798-2_16.
We present here a CRISPR-interference-based protocol to trigger prophage induction, even for non-inducible prophages. This method can also be used to cure the prophage from the bacterial host. The method is based on silencing of the phage's repressor transcription, thanks to CRISPR interference. Plasmid electroporation is used to bring the CRISPRi system into the bacteria, specifically on a plasmid carrying spacers targeting the prophage repressor. This method enables prophage induction and curation in a week or two with a high efficiency.
我们在此提出了一种基于 CRISPR 干扰的方法来触发噬菌体诱导,即使是非诱导性噬菌体也可以。这种方法也可以用于从细菌宿主中清除噬菌体。该方法基于 CRISPR 干扰来沉默噬菌体的阻遏物转录。通过电穿孔将 CRISPRi 系统导入细菌,特别是导入带有靶向噬菌体阻遏物间隔序列的质粒。这种方法可以在一到两周内高效地诱导和清除噬菌体。