Research Paper Volume 15, Issue 24 pp 14509—14552

Figure 2. Changes in IgG glycan composition upon targeting of selected GWAS loci associated with IgG galactosylation (HIVEP2, MANBA, TNFRSF13B and EEF1A1) in dCas9-VPR monoclonal cell lines. Samples containing non-targeting gRNAs served as controls. Changes in transcript levels are given as fold change values and changes in IgG phenotype are given as a relative change compared to control samples. (A) Manipulation of the HIVEP2 gene did not result in a statistically significant change in HIVEP2 transcript level, however, did induce a significant increase of digalactosylated structures (G2) with a concomitant decrease of agalactosylated IgG glycan structures (G0). (B) Targeting of MANBA by dCas9-VPR elevated transcription level of this gene which resulted in decrease of monogalactosylated IgG glycan structures (G1) (C) Targeting TNFRSF13B by dCas9-VPR resulted in ~ 400-fold increase of transcript levels and significant change of IgG agalactosylated IgG glycans (G0). (D) Successful upregulation of the EEF1A1 locus was followed by an increase of agalactosylated IgG glycans (G0) with a concomitant decrease of monogalactosylated IgG glycans (G1). Nominal p-value: *<0.05; **<0.01; ***<0.001; ns, not significant.