Figure 2 | Scientific Reports

Figure 2

From: Necessity of integrated genomic analysis to establish a designed knock-in mouse from CRISPR-Cas9-induced mutants

Figure 2

Analysis of Venus gene copy number using droplet digital PCR. (A) Oxytocin receptor (OXTR)-Venus knock-in heterozygous mice used as 1-copy positive controls. Schematic representation of the wild-type mouse OXTR genomic locus (Upper) and the genomic locus with Venus knock-in are shown (Lower). Exon 4 (E4) of the OXTR gene was used to normalize the Venus copy number as a 2-copy reference gene. The green and blue bars show specific probe sets for OXTR and Venus, respectively. (B) Reliable detection of the Venus gene copy number using genomic DNA from OXTR-Venus knock-in heterozygous mice generated by embryonic stem cells. Representative droplet plots of oxytocin receptor Venus/ + and + / + (Venus-positive droplets [Upper left], OXTR positive-droplet [Lower left]). Calculated Venus gene copy number in oxytocin receptor Venus/ + and + / + mice (right). (C) Calculated copy numbers of the Venus gene in the F0, F1, and F2 generations of the Prlhr-Venus knock-in mice. Black arrows show the parent–offspring relationships.

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