Fig. 3

Modeling CRISPR/Cas9-mediated DNA deletion using a Lox-STOP-Lox (LSL) reporter. a Schematic of the LSL cassette (STOP is 2.7 kb) of a reporter plasmid (LSL). Purple triangles indicate the LoxP sites recognized by sgLoxP. The asymmetric 8 bp sequence of LoxP is underlined. The red arrowhead indicates the Cas9 cutting site. The red arrows indicate the location and direction of forward and reverse primers, respectively. The ‘NAG’ PAM in the LoxP sequence is in bold. b 293T cells were co-transfected with 0.3 μg LSL and 0.5 μg sgLoxP and imaged 48 h later. c The level of luciferase bioluminescence was quantified. Error bars are the standard deviation (s.d., n = 3). d A PCR reaction-detected deletion. An arrowhead indicates the expected deletion band. e PCR samples were purified, TOPO cloned, and sequenced. Red nucleotides indicate indels. f Deep sequencing. Representative IGV images of two biological replicates. g Count of indels