Pvax1 plasmid vector

The type A outer surface protein C (OspC) gene of Borrelia burgdorferi B31 (GenBank accession #AAC66329.1) was designed to include the human tyrosinase signal peptide (MLLAVLYCLLWSFQTSAGHFPRA; GenBank accession #AH003020) at the N-terminus (30 (link), 31 (link)). The coding region was optimized for expression in mice and commercially synthesized in a pUC57 plasmid vector by Bio Basic Inc. (Markham, ON). The OspC gene containing the signal sequence was sub-cloned into a pVAX1 plasmid vector (ThermoFisher, Ottawa, ON), using NotI-HF and EcoRI-HF restriction enzymes (New England Biolabs, Whitby, ON). Large-scale amplifications of the pVAX1-OspC plasmid were generated using the QIAGEN Plasmid Giga Kit (Montreal, QC) according to the manufacturer’s instructions. The genetic sequences were validated by Sanger Sequencing prior to nanoparticle encapsulation.

mRNA-encoding firefly luciferase (mRNA-Luc) was purchased from TriLink Biotechnologies (San Diego, CA, USA) with the substitution of N1-Methylpseudouridine for uridine.
To generate the pDNA-encoding firefly luciferase gene (pVAX1-Luc), the Luc gene from the pcDNA3-Luc plasmid (Addgene, Watertown, MA, USA) was cloned into the pVAX1 plasmid vector (ThermoFisher, Ottawa, ON, Canada) using HindIII-HF and XbaI restriction enzymes (New England Biolabs, Whitby, ON, Canada). pVAX1-Luc was transformed into NEB^® 10-beta Competent E. coli (High Efficiency) (New England Biolabs, Ipswich, MA, USA)according to the manufacturer’s protocol). Large-scale amplifications of pVAX1-Luc were generated using the EndoFree QIAGEN Plasmid Giga Kit (Montreal, QC, Canada) according to the manufacturer’s instructions.