Fig. 1

Schematic diagram of BgAIF gene structure, CRISPR/Cas9 vector and expression in Bge cell. a Gene structure of B. glabrata allograft inflammatory factor (BgAIF), accession number BGLB005061 and gene editing target locus (red box) on exon 4. BgAIF gene composed of 5 exons and 4 introns. The green arrows indicate the location of primers flanking expected double-strand breaks (DSB) which were used in PCR to generate the on-target amplicon for INDELs estimation. b Map of the pCas-BgAIFx4 vector which includes the Pol III-dependent mammalian U6 gene promoter (red arrow) to drive transcription of the guide RNA targeting exon 4 of BgAIF gene (red arrow) and the CMV promoter to drive expression of the S. pyogenes Cas9 nuclease (blue arrow). Primer pairs specific for the guide RNA and for Cas9 are indicated (green arrows). c Expression of Cas9 and of BgActin (as the reference gene) transcripts as established by semi-quantitative RT-PCR in pCas-BgAIF-transfected (right) and control (left) Bge cells from days one to nine following transfection. The amplicons of the expected sizes are as indicated: 23 bp for Cas9 and 214 bp for BgActin. All RNA samples were positive for the BgActin reference gene; the 214 bp band