[ODP-221] Generation of phagemid-based CRISPR-Cas13 antimicrobials against MRSA
Various antibiotic-resistant bacteria have been emerging worldwide, creating a need for the development of new effective antimicrobials. We previously reported development of programmable, sequence-specific antimicrobials by loading CRISPR-Cas13 genes onto bacteriophage capsid taking advantage of hijacking mechanism of Staphylococcus aureus pathogenicity island (SaPI). SaPI can be packaged into phage capsid, however, type of phage that accept SaPI is limited. In this study, we developed a phagemid-based packaging system to load CRISPR-Cas13 genes into phage capsid in order to apply the packaging system to various phages. The resulting phage capsids carrying CRISPR-Cas13 phagemid (denoted as antimicrobial Capsid, AB-Capsid) were tested for their ability to kill bacteria in target gene sequence-specific manner. As an example, generated mecA AB-Capsid carrying CRISPR-Cas13 designed to target mecA gene showed killing effect against MRSA but not MSSA. Further study to develop and improve the capsid packaging technology capable of delivering CRISPR-Cas13 genes to a wide range of bacteria species is undergoing.