Figure 2
From: Baf53a is involved in survival of mouse ES cells, which can be compensated by Baf53b
Strategy for generation of tetracycline (Tet)-inducible Baf53a conditional knockout (cKO) ES cells. (A) The targeting site of Baf53a. The targeting site (5′-GTT GAT TTC CCC ACG GCT AT-3′) of the Baf53a gene was shown. The target sequences are located on Exon 3 of the gene. (B) Alignment of the target sites of wild-type Baf53a and CRISPR/Cas9 resistant Baf53a (Baf53aR). The targeting site of the Baf53a cDNA was changed to 5′-GTT GAc TTt CCt ACc GCc AT-3,’ which encodes an identical amino acid sequence. (C) Schematic view of Tet-inducible Baf53a cKO ES cells. Expression of exogenous Baf53aR was induced in the absence of Tet and repressed in the presence of Tet. Both alleles of the Baf53a gene were mutated by the CRISPR/Cas9 system. (D) Sequences of deleted alleles. Genomic DNA of the deleted regions was sequenced. One allele contained a 7 bp deletion and the other contained a 23 bp deletion.
