HARMONIUM, INC.

The 2014 Ebola outbreak in West Africa highlighted the need for effective vaccines. The rVSV-ZEBOV vaccine, developed through extensive trials, marked a turning point with near-perfect efficacy. Now, Ervebo is crucial in combating Ebola, especially the Zaire strain, but challenges like vaccine hesitancy and supply issues persist.

Research adheres to ethical regulations, with animal experiments approved by UC Berkeley's ACUC. Plasmid construction involves pCold vector-based Cas protein expression, with specific tags and cleavage sites. Nucleic acids and lipid materials are prepared and quality-controlled. IVT sgRNA production and directed evolution of GeoCas9 are detailed. Protein expression and purification methods are described, including use of Ni-NTA and heparin affinity columns. Cell lines and culture conditions are specified for various experiments. RNP assembly, genome editing, flow cytometry, and LNP delivery are outlined. In vivo genome editing and immunogenicity assessment methods are also detailed.