Fig 4. CRISPR/Cas9 mediated indels in the ECFP gene of PUBB2 P61 x HWE hybrids.

(A) Sanger-sequencing trace data showing the regions of the indels in mutants P41, P49, P55, and F82. Arrows indicate the cleavage site; the sequence of guide RNA sg35 is shadowed. (B) Nucleotide sequence alignments showing indels. The ECFP nucleotide sequence is shown at the top with the sg35 target site in bold-type and the PAM sequence in red. Deletions are shown as dashes. cDNA fragments spanning each target site were PCR-amplified using genomic DNA from pooled larvae as template and cloned into the pCR4-TOPO TA vector. At least 10 cDNA clones per PCR amplicon were sequenced.