Fig. 2 | Scientific Reports

Fig. 2

From: Double-strand break-free and transgene-free genome editing in the microalga Nannochloropsis oceanica using removable vectors containing the CRISPR base editing system

Fig. 2

Analysis of base substitution at the NoNR target site in ArBE-induced strains (A) Target site sequences of the endogenous nitrate reductase gene, NoNR, for ArBE vectors. Underlined “GT” and “AG” represent the junction sites of exon and intron in the NoNR gene. PAMs: Protospacer adjacent motifs. (B) Chromatogram data from ABI sequencing of WT genomic DNAs which indicated “WT” or mixed genomic DNA which indicated “BE” and extracted from colonies that emerged on selective plates containing ArBE vectors targeting NoNR. Red boxes show the positions of the intron–exon junctions. Arrows indicate the position of gRNAs. (C) Data analysis of base editing at sgRNA base positions using chromatogram data of ABI sequencing by EditR. Base positions indicate the distance of sgRNA spacer bases from the PAMs. (D) Chromatogram data of ABI sequencing using single colonies that emerged on selective plates of ArBE vectors for NoNR. Underbars show the position of intron–exon junctions. The white text on the black background indicates the bases substituted by ArBE vectors.

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