Figure 3
From: A CRISPR-Cas9 based shuffle system for endogenous histone H3 and H4 combinatorial mutagenesis
Generation of histone H3 single and triple mutants through a single transformation. (A) Graphic representation of CRISPR-Cas9 based mutagenesis on histone H3. The transformation was performed on synH3 strain with 2 targeting gRNAs and donors harboring mutation sites and HR regions. The single piece donors were generated through fusion PCR. (B) The mutagenesis efficiencies for generating histone H3 single and triple mutant through transformation with different formats of donors. The repairing donors can be prepared by either multiple donor fragments with flanking HR arms for mutation or two single pieces of donors containing mutation sites. The data of bar charts represent mean averages of over-all mutagenesis efficiencies of 3 biological replicates with each randomly picked 8 colonies. The error bars indicate the standard deviations of 3 biological replicates. (C) The representation of sequencing results of both Histone H3 K4R–K36R–K79R at endogenous loci. The sequence alignments were performed by SnapGene software.
