Fig. 5

CRISPR/Cas9-mediated editing of the EcN genome with MPER insert. A schematic representation of the development of the EcN-MPER clones using CRISPR/Cas9 according to [29] with modifications. (1) Transformation of EcN with plasmids and the template oligo; (2) Nuclease Cas9 is guided by gRNA to create a site-specific break in bacterial gDNA; (3) OmpF-MPER oligonucleotide is used as the template for gDNA repair through homologous recombination; (4) PCR and sequencing to confirm the insertion of MPER in the desired position inside ompF gene