Reprogrammable bacterial nanosyringes for RNA and gene editors delivery

The engineered Photorhabdus virulence cassette (PVC) has not yet achieved the RNA packaging and injection delivery, although it can accurately deliver diverse proteins. Fortunately, RNA-binding proteins have a high affinity for binding to specific sequences and are excellent anchoring components between RNA and the inner tube of PVC. By combining the U1A RNA-binding domain (U1A RBD) and PVC, we developed a convenient, RNA non-degradable, and universal PVC-based RNA delivery tool, establishing DART (PVC Docker-based All-purpose RNA Injection Delivery Tool). DART successfully achieved the delivery of diverse RNAs, including Pepper RNA, gRNA, siRNA, and miRNA. We also demonstrated that the DART enabled effective knockout of GFP in vivo and in vitro. Our DART system provided a powerful platform for RNA and gene editing therapeutics in vivo.