Fig. 3
From: Versatile generation of precise gene edits in bovines using SEGCPN
Generation of blastocysts and fetuses with an 11-bp MSTN deletion using SEGCPN. a Procedure for generating blastocysts and fetuses with an 11-bp MSTN deletion using SEGCPN. b Identification of MSTN 11-bp deletion edited cell clones by PCR. M, 1-kb DNA ladder; C1–C10, MSTN-edited clones; P, donor vector; WT, wild-type cells; H2O, negative control. c Identification of MSTN 11-bp deletion-edited blastocysts by PCR. M, 100-bp DNA ladder; embryo+/−, heterozygous MSTN-edited embryos; embryo−/−, homozygous MSTN-edited embryos; WT, wild-type embryos; H2O, negative control. d Sanger sequencing chromatograms of DNA from WT and MSTN mutant blastocysts. The red arrowhead indicates the 11-bp deletion. e Alignments of mutant sequences from the TA-cloning sequencing of a single embryo. The 11-bp deletion site is shown in red. The column on the right indicates the frequencies of mutant alleles. WT, wild-type embryos. f Identification of mosaicism in a homozygous MSTN−/− 11-bp deletion edited fetus. P4/P5, identification of MSTN−/− gene editing in various fetal tissues. Neo-F/R, the marker gene in the ESSEE detected in various fetal tissues. Cell−/−, homozygous positive gene-edited clone; lanes 3–9, various tissues of the MSTN−/− fetus; WT, wild-type tissue; P, donor vector; H2O, negative control