Figure 3 | Scientific Reports

Figure 3

From: A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse

Figure 3

c-Maf/MafB double cKO via sgRNAs-LSL-Cas9 system. (a) Schematic of transgenic vector sgRNAsc-Maf/MafB-UbC-LSL-Cas9. Three U6 promoters (green arrowheads) driven c-Maf sgRNAs (blue blocks) and three U6 promoters driven MafB sgRNAs (green blocks) were arranged alternately. Other functional regions are combined as those in the sgRNAsEGFP-UbC-LSL-Cas9 transgenic vector (Fig. 1a). (b) Photograph of sgRNAs c-Maf/MafB-UbC-LSL-Cas9 mice. Ctrl, transgenic mouse without Cre. LysM-Cre, transgenic mouse express lysM-Cre. (c) Western blots of c-Maf/MafB double cKO macrophages (paired t-test, c-Maf, ***p = 0.003, n = 3. MafB, ***p = 0.0009, n = 3). (d) T7EN1 analysis of c-Maf and MafB mutations in the macrophages of a sgRNAs c-Maf/MafB-LSL-Cas9; LysM-Cre (LysM-Cre) mouse. The control (Ctrl) was a sgRNAs c-Maf/MafB-LSL-Cas9 mouse without Cre recombinase. M, marker. (e) c-Maf mutation mode of the Cre-expressing macrophages of the transgenic mouse. (f) MafB mutation mode of the Cre-expressing macrophages of the transgenic mouse.+, insertion.

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