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. 2021 Nov 16;2(4):100965. doi: 10.1016/j.xpro.2021.100965

Figure 5.

Figure 5

Insertion of loxP sequence using ssODN

(A) Schematic to insert a loxP sequence at the DMD gene locus in 1383D2 iPSCs. Successful insertion of a loxP site generates an extra XmnI restriction enzyme site, which can be detected by DNA electrophoresis.

(B) Cas9, gRNA, and ssODN were electroporated into 1383D2 iPSCs and genomic DNA was extracted for PCR amplification. TapeStation analysis of the PCR products with or without XmnI digestion. With the RNP and ssODN electroporation condition, extra bands with expected sizes (229 bp and 67 bp) were observed, suggesting the loxP sequence is inserted at the target site with around 24% of efficiency. This figure is adapted from Kagita et al. (2021).