Extended Data Fig. 2: Off-target analysis of generated CRISPR clones. | Nature Genetics

Extended Data Fig. 2: Off-target analysis of generated CRISPR clones.

From: Lysine 4 of histone H3.3 is required for embryonic stem cell differentiation, histone enrichment at regulatory regions and transcription accuracy

Extended Data Fig. 2: Off-target analysis of generated CRISPR clones.

a, Three predicted off-target sites for CRISPR guides used to introduce K4A and K36A mutations were amplified by PCR and analyzed by Sanger-Sequencing to confirm the absence of indels. b-d, Off-target analysis of two controls in b, three H3.3K4A in c, or three H3.3K36A in d, mutant cell lines to confirm their genomic integrity after CRISPR targeting. log2(FoldChanges) of gene expression in CRISPR clones compared to unmodified wild type ESCs were determined by DESeq2 and plotted over chromosome position. Lines indicate CIRSPR cleavage site inside H3f3a gene (yellow) and H3f3b gene (blue). Loss or duplication of a chromosome part can be detected by coordinated up- or down-regulation of close-by genes. All clones included in this study do not have detectable chromosome deletion/duplications.

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