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Fig. 5 | BMC Genomics

Fig. 5

From: Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ

Fig. 5

Potential window of MMEJ-dependent base replacement revealed by sequence alignment of on- and off-target knock-ins. a Sequence alignment of upstream and downstream regions of the DSB site. OT1–OT14 represents off-target sites, related to Additional file 1: Table S1. Roman numbers shown at the right side of each set of sequences indicate the allele types, related to Fig. 4d, e and Additional file 1: Figure S15, S16 and Table S1. Black triangles indicate the DSB site. Black boxes indicate the CRISPR target sequence. Red and blue underlines indicate the left and right microhomologies, respectively. Black underlines indicate mutated sequences found in sequenced knock-in junctions. Blue letters indicate the polymorphisms found in sequenced knock-in junctions, related to Fig. 4d, e. Red letters indicate the mutations not found in the sequenced knock-in junctions. Asterisks indicate positions with complete base conservation among on- and off-target sites. The positions of bases replaced and not replaced with the donor sequence were masked in red and blue, respectively. The upstream sequences of OT2, OT3, OT4, OT5, OT7, and OT8 and the downstream sequences of OT5, OT6, and OT8 were identical, respectively (Additional file 1: Figure S17). b Schematic representation of potential MMEJ window. Red and blue boxes indicate the left and right microhomologies, respectively

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