Extended Data Figure 10: Generation of Gbpchr3 mice by CRISPR/Cas9-mediated genome editing and analyses of its functional connection with ipaH7.8.
From: Ubiquitination and degradation of GBPs by a Shigella effector to suppress host defence
a, The Gbp locus on mouse chromosome 3 and the genomic RNA targeting strategy for generating the knockout mice. b, Genome typing of founder mice by PCR. +/+, +/− and −/− denote mice with wild-type Gbp locus, one copy of the Gbp locus deleted and both copies of the Gbp locus deleted, respectively. Chromosomal location of the four PCR primers used for genome typing are shown in a. c, Junction sequences of two types of Gbpchr3 deletion alleles, determined by DNA sequencing of the PCR products in b. Mice used in this study are homologous for one of the two out-of-frame alleles or heterozygous for the two alleles. d–g, Wild-type or Gbpchr3 mice (C57BL/6 background) were infected with S. flexneri ΔipaH7.8 intravenously (d, e) or intraperitoneally (f, g) at MOI of 2 × 106 or 4 × 106, respectively. Mouse survival analysis (d, f) was performed in GraphPad Prism 5. Bacterial loads 1 day after infection (e, g) are expressed as log10 colony-forming units per gram of liver or spleen tissue; mean values are also shown. Sample sizes (biologically independent animals): n = 10 for both groups in d, n = 7 for both groups in e, n = 12 for both groups in f, n = 7 for WT/ΔipaH7.8 and n = 6 for Gbpchr3/ΔipaH7.8 in g. Data shown are representative of three (b) or two (d–g) independent experiments.
