Fig. 4: Frequencies of delinver mutations are positively correlated with those of deletion mutations. | Nature Communications

Fig. 4: Frequencies of delinver mutations are positively correlated with those of deletion mutations.

From: Hidden prevalence of deletion-inversion bi-alleles in CRISPR-mediated deletions of tandemly arrayed genes in plants

Fig. 4

a Diagram of the AT5G45240/AtRPS4/AtRRS1 locus. Three gRNA-Ls (L1 to L3) targeting AT5G45240 and ten gRNA-Rs (R1 to R10) targeting AtRRS1 were designed for testing the frequencies of delinver mutations mediated by different gRNA-L/gRNA-R pairs. gRNAs targeting to antisense or sense strands are indicated by leftwards and rightwards arrows, respectively. Note that the gRNAs R2-R6 were targeted to the same region of AtRRS1. Colored letters mark PAMs. L-Fw2/R-Fw2 and L-Rev2/R-Rev2 are examples of co-aligned primer pairs, which were used for detecting genomic inversions between gRNA-L2 and gRNA-R9 induced cut sites. b Representative Sanger sequencing results of PCR amplicons using co-aligned primers validated the inversion between gRNA-L2 and gRNA-R9 induced breakpoints in the L2 + R9 mutant line #7. Black bold letters mark PAMs and target sequences of gRNAs are underlined. c Frequencies of delinver mutations are positively correlated with those of deletion mutations. d Frequencies of delinver mutations are unrelated to the deletion sizes. In c and d each dot represents the data of one gRNA pair and the Pearson correlation was calculated using the GraphPad Prism algorithm.

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