Characterization of Photorhabdus Virulence Cassette as a causative agent in the emerging pathogen Photorhabdus asymbiotica

N-terminal signal peptides facilitate the engineering of PVC complex as a potent protein delivery system

“…S1A). Two effectors, photorhabdus necrotizing factor (Pnf) and photorhabdus dNTP pyrophosphatase 1 (Pdp1), can be loaded and translocated by PVC-V complex (hereafter PVC unless indicated) for cytotoxicity ( 13 ). Through comparison between the PVC effectors and their homologs, we have found that a typical extra N-terminal fragment can be defined within the protein sequences of PVC effectors (Fig.…”

“…2B). Pnf and Pdp1 effectors were previously proven to be injected by the PVC complex into J774 murine macrophages ( 13 ). To test the effects of reporters after PVC delivery, we treated J774 with relative PVC solutions, and relevant measurements were carried out.…”

“…For the construction of pCNM3 and pBR60 expressing LysR regulator gene, detailed procedures can be referred previously ( 12 , 13 ). To generate pCNM12 plasmid producing both PVC particle and its effectors, the genomic DNA fragment (PAU_03353 to PAU_03332) was cleaved using Cas9-Assisted Targeting of Chromosome Segments as described ( 49 ) and was ligated into the pRK404 for expression.…”

Potential of Mitochondrial Genome Editing for Human Fertility Health

“…Practically, this RP signal has been used to import wild-type tRNAs efficiently into human mitochondria ( Wang et al, 2012 ). A recent study showed that the mtDNA could be targeted with engineered CRISPR/Cas9 to assess the mutagenesis outcome of nuclease-generated DSBs in human cells ( Wang et al, 2021 ). Site-specific DSBs were generated via Staphylococcus aureus Cas9 fused with the mitochondrial targeting sequence of COX8A in mtDNA, providing novel potential for the treatment of mitochondrial diseases with MGE ( Wang et al, 2021 ).…”

“…A recent study showed that the mtDNA could be targeted with engineered CRISPR/Cas9 to assess the mutagenesis outcome of nuclease-generated DSBs in human cells ( Wang et al, 2021 ). Site-specific DSBs were generated via Staphylococcus aureus Cas9 fused with the mitochondrial targeting sequence of COX8A in mtDNA, providing novel potential for the treatment of mitochondrial diseases with MGE ( Wang et al, 2021 ). Moreover, appending different signal sequences to gRNAs has recently been discussed, providing the first evidence that this is indeed a viable strategy to develop mito-CRISPRs ( Anton et al, 2020 ).…”